gracilaria chilensis total rna  (Qiagen)


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    RNeasy Plant Mini Kit
    Description:
    For purification of total RNA from plants and fungi Kit contents Qiagen RNeasy Plant Mini Kit 20 preps 10 to 100mg Sample 30 to 100L Elution Volume Plant Sample Total RNA Purification Spin Column Format Silica Technology Ideal for Northern Dot and Slot Blotting End point RT PCR Quantitative Real time RT PCR Array Analysis Includes 20 RNeasy Mini Spin Columns 20 QIAshredder Mini Spin Columns Collection Tubes 1 5mL and 2mL RNase free Reagents and Buffers Benefits High quality total RNA in 30 minutes No phenol chloroform extraction No CsCl gradients no LiCl or ethanol precipitation Excellent recovery of RNA Ready to use RNA for any downstream applicatio
    Catalog Number:
    74903
    Price:
    155
    Category:
    RNeasy Plant Mini Kit
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    Structured Review

    Qiagen gracilaria chilensis total rna
    RNeasy Plant Mini Kit
    For purification of total RNA from plants and fungi Kit contents Qiagen RNeasy Plant Mini Kit 20 preps 10 to 100mg Sample 30 to 100L Elution Volume Plant Sample Total RNA Purification Spin Column Format Silica Technology Ideal for Northern Dot and Slot Blotting End point RT PCR Quantitative Real time RT PCR Array Analysis Includes 20 RNeasy Mini Spin Columns 20 QIAshredder Mini Spin Columns Collection Tubes 1 5mL and 2mL RNase free Reagents and Buffers Benefits High quality total RNA in 30 minutes No phenol chloroform extraction No CsCl gradients no LiCl or ethanol precipitation Excellent recovery of RNA Ready to use RNA for any downstream applicatio
    https://www.bioz.com/result/gracilaria chilensis total rna/product/Qiagen
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    gracilaria chilensis total rna - by Bioz Stars, 2020-04
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    Images

    1) Product Images from "Molecular and functional characterization of ferredoxin NADP(H) oxidoreductase from Gracilaria chilensis and its complex with ferredoxin"

    Article Title: Molecular and functional characterization of ferredoxin NADP(H) oxidoreductase from Gracilaria chilensis and its complex with ferredoxin

    Journal: Biological Research

    doi: 10.1186/s40659-017-0144-5

    a Nucleotide sequence of the gene identified for FNR of Gracilaria chilensis . Nucleotides 1–135 (bold letters) codify for a chloroplastide transit signal. b Translated sequence of the G. ch mature protein, from amino acid 1. A sequence alignment with FNR from Zea mays, Pyropia yezoensis, Chondrus crispus, Synechococcus sp. and Anabaena sp .* is also shown. In Synechococcus , the sequence for a PBS binding third domain is shown (residues – 1 to − 80). The NADP and FAD binding domains are indicated by arrows, the conserved residues for the binding of co-factor and substrate are also shown in blue squares and red squares for FAD and NADP binding residues respectively.Amino acid residues involved in the interaction with ferredoxin are also shown. *The third domain for Anabaena sp. is not shown for the clarity of the alignment. Only the common two domains present in the structural complex (1ewy) are shown
    Figure Legend Snippet: a Nucleotide sequence of the gene identified for FNR of Gracilaria chilensis . Nucleotides 1–135 (bold letters) codify for a chloroplastide transit signal. b Translated sequence of the G. ch mature protein, from amino acid 1. A sequence alignment with FNR from Zea mays, Pyropia yezoensis, Chondrus crispus, Synechococcus sp. and Anabaena sp .* is also shown. In Synechococcus , the sequence for a PBS binding third domain is shown (residues – 1 to − 80). The NADP and FAD binding domains are indicated by arrows, the conserved residues for the binding of co-factor and substrate are also shown in blue squares and red squares for FAD and NADP binding residues respectively.Amino acid residues involved in the interaction with ferredoxin are also shown. *The third domain for Anabaena sp. is not shown for the clarity of the alignment. Only the common two domains present in the structural complex (1ewy) are shown

    Techniques Used: Sequencing, Binding Assay

    a Comparative model of ferredoxin NADP + reductase from Gracilaria chilensis. Secondary structures are shown by arrows (β strands) and cylinders (α helices); FAD is shown as orange sticks and NADP as purple sticks. b Close up of the residues involved in the NADP + binding site, c close up of the residues involved in the FAD binding site. d Docking model of FNR (grey surface), showing the two co-substrates, NADP + and ferredoxin, the co-factor FAD and the 2[FeS] cluster. e Close-up of the complex showing residues of the interface included in the text
    Figure Legend Snippet: a Comparative model of ferredoxin NADP + reductase from Gracilaria chilensis. Secondary structures are shown by arrows (β strands) and cylinders (α helices); FAD is shown as orange sticks and NADP as purple sticks. b Close up of the residues involved in the NADP + binding site, c close up of the residues involved in the FAD binding site. d Docking model of FNR (grey surface), showing the two co-substrates, NADP + and ferredoxin, the co-factor FAD and the 2[FeS] cluster. e Close-up of the complex showing residues of the interface included in the text

    Techniques Used: Binding Assay

    Unrooted phylogenetic tree built using the sequences of Gracilaria chilensis (Rhodophyta, eukaryote) Chondrus crispus (Rhodophyta, eukaryote), Pyropia yezoensis (Rhodophyta, eukaryote), Cyanidium caldarium (Rhodophyta, eukaryote), Cyanophora paradoxa (Glaucophyta, eukaryote), Thermosynechococcus elongates (Cyanobacteria, prokaryote), Fremyella diplosiphon (Cyanobacteria, prokaryote), Anabaena variabilis (Cyanobacteria, prokaryote), Synechococcus elongates (Cyanobacteria, prokaryote), Arthrospira platensis (Cyanobacteria, prokaryote), Chlamydomonas reinhardtii (Chlorophyta, eukaryote), Nicotiana tabacum (Magnoliophyta, eukaryote), Pisum sativum (Anthophyta, eukaryote), Arabidopsis thaliana (Tracheophyta, eukaryote), Zea mays (Magnoliophyta, eukaryote), Oryza sativa (Magnoliophyta, eukaryote), and Spinacia oleracea (Streptophyta, eukaryote). Letters in parenthesis indicate leaf (L) or root (R) isoforms
    Figure Legend Snippet: Unrooted phylogenetic tree built using the sequences of Gracilaria chilensis (Rhodophyta, eukaryote) Chondrus crispus (Rhodophyta, eukaryote), Pyropia yezoensis (Rhodophyta, eukaryote), Cyanidium caldarium (Rhodophyta, eukaryote), Cyanophora paradoxa (Glaucophyta, eukaryote), Thermosynechococcus elongates (Cyanobacteria, prokaryote), Fremyella diplosiphon (Cyanobacteria, prokaryote), Anabaena variabilis (Cyanobacteria, prokaryote), Synechococcus elongates (Cyanobacteria, prokaryote), Arthrospira platensis (Cyanobacteria, prokaryote), Chlamydomonas reinhardtii (Chlorophyta, eukaryote), Nicotiana tabacum (Magnoliophyta, eukaryote), Pisum sativum (Anthophyta, eukaryote), Arabidopsis thaliana (Tracheophyta, eukaryote), Zea mays (Magnoliophyta, eukaryote), Oryza sativa (Magnoliophyta, eukaryote), and Spinacia oleracea (Streptophyta, eukaryote). Letters in parenthesis indicate leaf (L) or root (R) isoforms

    Techniques Used:

    Sequences of ferredoxins as detected in the transcriptome of Gracilaria chilensis. a Alignment of the FdS with FdL. b Sequence alignment among FdS and ferredoxins from Z. mays (1gaq) and Anabaena sp. (1ewy). Only the identities are shown as (*), the cysteines involved in the binding of the FeS center are displayed in red
    Figure Legend Snippet: Sequences of ferredoxins as detected in the transcriptome of Gracilaria chilensis. a Alignment of the FdS with FdL. b Sequence alignment among FdS and ferredoxins from Z. mays (1gaq) and Anabaena sp. (1ewy). Only the identities are shown as (*), the cysteines involved in the binding of the FeS center are displayed in red

    Techniques Used: Sequencing, Binding Assay

    2) Product Images from "Multiplex Detection of Aspergillus fumigatus Mycoviruses"

    Article Title: Multiplex Detection of Aspergillus fumigatus Mycoviruses

    Journal: Viruses

    doi: 10.3390/v10050247

    Conventional PCR to check amplicon size prior to multiplex PCR. Amplicon sizes were checked on two percent agarose gel prior to performing multiplex PCR for A. fumigatus dsRNA mycoviruses. AfuCV, AfuPV-1 and AfuTmV-1 dsRNAs were extracted using LiCl extraction (Lanes 2, 4 and 6, respectively) and RNeasy Plant Mini kit (Lanes 3, 5 and 7, respectively) and used as templates for amplification with AfuCV, AfuPV-1 and AfuTmV-1 primers (2–3, 4–5, 6–7, respectively). Hyperladder-I was used as a marker to estimate the size of the amplicons (Lane 1).
    Figure Legend Snippet: Conventional PCR to check amplicon size prior to multiplex PCR. Amplicon sizes were checked on two percent agarose gel prior to performing multiplex PCR for A. fumigatus dsRNA mycoviruses. AfuCV, AfuPV-1 and AfuTmV-1 dsRNAs were extracted using LiCl extraction (Lanes 2, 4 and 6, respectively) and RNeasy Plant Mini kit (Lanes 3, 5 and 7, respectively) and used as templates for amplification with AfuCV, AfuPV-1 and AfuTmV-1 primers (2–3, 4–5, 6–7, respectively). Hyperladder-I was used as a marker to estimate the size of the amplicons (Lane 1).

    Techniques Used: Polymerase Chain Reaction, Amplification, Multiplex Assay, Agarose Gel Electrophoresis, Marker

    3) Product Images from "Filter paper-based spin column method for cost-efficient DNA or RNA purification"

    Article Title: Filter paper-based spin column method for cost-efficient DNA or RNA purification

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0203011

    The efficiency of filter paper for purification of nucleic acids from various sources using respective Qiagen kits. (A) Tomato genomic DNAs purified using Qiagen DNeasy plant mini kit. (B) Tomato total RNAs purified using Qiagen RNeasy plant mini kit. (C) PCR products of a GUS fragment purified using Qiagen QIAquick PCR purification kit. (D) PCR products of GUS fragment recovered from an agarose gel using a Qiagen QIAquick gel extraction kit. (E) pUC -19 plasmid DNAs purified using a Qiagen QIAprep spin miniprep kit. For each panel, from left to right are (Q) nucleic acid purified in experiments using original Qiagen spin column, (G) reassembled spin column using two layers of Whatman glass microfiber filters (Grade GF/F), and (P) reassembled spin column using two layers of Whatman qualitative filter paper, (Grade 3) respectively. Upper panel is quantification data based on three experimental replicates normalized according to performance of the Qiagen kit; lower panel is an image of agarose gel electrophoresis for the same volume of purified nucleic acids.
    Figure Legend Snippet: The efficiency of filter paper for purification of nucleic acids from various sources using respective Qiagen kits. (A) Tomato genomic DNAs purified using Qiagen DNeasy plant mini kit. (B) Tomato total RNAs purified using Qiagen RNeasy plant mini kit. (C) PCR products of a GUS fragment purified using Qiagen QIAquick PCR purification kit. (D) PCR products of GUS fragment recovered from an agarose gel using a Qiagen QIAquick gel extraction kit. (E) pUC -19 plasmid DNAs purified using a Qiagen QIAprep spin miniprep kit. For each panel, from left to right are (Q) nucleic acid purified in experiments using original Qiagen spin column, (G) reassembled spin column using two layers of Whatman glass microfiber filters (Grade GF/F), and (P) reassembled spin column using two layers of Whatman qualitative filter paper, (Grade 3) respectively. Upper panel is quantification data based on three experimental replicates normalized according to performance of the Qiagen kit; lower panel is an image of agarose gel electrophoresis for the same volume of purified nucleic acids.

    Techniques Used: Purification, Polymerase Chain Reaction, Agarose Gel Electrophoresis, Gel Extraction, Plasmid Preparation

    Evaluation of purification of tobacco genomic DNA and total RNA using filter paper-based spin columns with respective Qiagen kit buffers and homemade buffers. (A) Agarose gel electrophoresis for 2.5 μl tobacco genomic DNAs elution from purification experiments using Qiagen DNeasy plant mini kit buffers with Qiagen original spin column (Lane Q/Q), filter paper recharged used spin column (Lane Q/R) and filter paper-based homemade spin column (Lane Q/H*), followed by tobacco genomic DNAs purified using homemade buffer with Qiagen original spin column (Lane H/Q), filter paper recharged used spin column (Lane H/R) and filter paper-based homemade spin column (Lane H/H*). (B) UV spectrum curve of tobacco DNAs purified using Qiagen kit (Q/Q, black curve), filter paper recharged spin columns with Qiagen kit buffers (Q/R, blue curve) or homemade buffers (H/R, red curve) from the same amount leaf tissue. Y-axis is UV absorbance, and X-axis is wavelength (nM). (C) Amplification plots for three duplicated qPCR reactions contain 20 ng DNA purified using Qiagen kit (Q/Q, Blue curves) or DNA purified from filter paper recharged spin column with homemade buffer (H/R, Red curves) respectively. The x-axis is PCR cycle numbers, Y-axis is the level of SYBR fluorescence, and the green line is an arbitrary threshold to determine the Cq value (the fractional cycle number at which amplification curve meet threshold level). (D) MOPS-formaldehyde denaturing agarose gel electrophoresis separated 5 μl RNA purified using Qiagen RNeasy plant mini kit buffers with a Qiagen original spin column (Lane Q/Q), filter paper recharged used spin column (Lane Q/R) and homemade filter paper-based spin column (Lane Q/H*), followed total tobacco RNAs purified by using homemade buffer with Qiagen original spin column (Lane H/Q), filter paper recharged used spin column (Lane H/R) and filter paper-based homemade spin column (Lane H/H*). (E) UV spectrum of tobacco total RNA purified using Qiagen kit (Q/Q, black curve), filter paper recharged spin column with Qiagen RNeasy plant mini kit buffers (Q/R, blue curve) or homemade buffers (H/R, red curve). Y-axis is UV absorbance, and the X-axis is wavelength. (F) Amplification plots of three duplicated qRT-PCR reactions for 2.5 ng RNA purified using Qiagen kit (Q/Q, Blue curves) or RNA purified using filter paper recharged spin column with homemade buffer (H/R, Red curves) respectively. Note: * The starting material amount is 100 mg tobacco leaf tissue for experiments using a Qiagen spin column or filter paper recharged spin column, and half amount of plant sample (50 mg) used for homemade spin column purification. All DNAs or RNAs were eluted using 100 ul elution solution.
    Figure Legend Snippet: Evaluation of purification of tobacco genomic DNA and total RNA using filter paper-based spin columns with respective Qiagen kit buffers and homemade buffers. (A) Agarose gel electrophoresis for 2.5 μl tobacco genomic DNAs elution from purification experiments using Qiagen DNeasy plant mini kit buffers with Qiagen original spin column (Lane Q/Q), filter paper recharged used spin column (Lane Q/R) and filter paper-based homemade spin column (Lane Q/H*), followed by tobacco genomic DNAs purified using homemade buffer with Qiagen original spin column (Lane H/Q), filter paper recharged used spin column (Lane H/R) and filter paper-based homemade spin column (Lane H/H*). (B) UV spectrum curve of tobacco DNAs purified using Qiagen kit (Q/Q, black curve), filter paper recharged spin columns with Qiagen kit buffers (Q/R, blue curve) or homemade buffers (H/R, red curve) from the same amount leaf tissue. Y-axis is UV absorbance, and X-axis is wavelength (nM). (C) Amplification plots for three duplicated qPCR reactions contain 20 ng DNA purified using Qiagen kit (Q/Q, Blue curves) or DNA purified from filter paper recharged spin column with homemade buffer (H/R, Red curves) respectively. The x-axis is PCR cycle numbers, Y-axis is the level of SYBR fluorescence, and the green line is an arbitrary threshold to determine the Cq value (the fractional cycle number at which amplification curve meet threshold level). (D) MOPS-formaldehyde denaturing agarose gel electrophoresis separated 5 μl RNA purified using Qiagen RNeasy plant mini kit buffers with a Qiagen original spin column (Lane Q/Q), filter paper recharged used spin column (Lane Q/R) and homemade filter paper-based spin column (Lane Q/H*), followed total tobacco RNAs purified by using homemade buffer with Qiagen original spin column (Lane H/Q), filter paper recharged used spin column (Lane H/R) and filter paper-based homemade spin column (Lane H/H*). (E) UV spectrum of tobacco total RNA purified using Qiagen kit (Q/Q, black curve), filter paper recharged spin column with Qiagen RNeasy plant mini kit buffers (Q/R, blue curve) or homemade buffers (H/R, red curve). Y-axis is UV absorbance, and the X-axis is wavelength. (F) Amplification plots of three duplicated qRT-PCR reactions for 2.5 ng RNA purified using Qiagen kit (Q/Q, Blue curves) or RNA purified using filter paper recharged spin column with homemade buffer (H/R, Red curves) respectively. Note: * The starting material amount is 100 mg tobacco leaf tissue for experiments using a Qiagen spin column or filter paper recharged spin column, and half amount of plant sample (50 mg) used for homemade spin column purification. All DNAs or RNAs were eluted using 100 ul elution solution.

    Techniques Used: Purification, Agarose Gel Electrophoresis, Amplification, Real-time Polymerase Chain Reaction, Polymerase Chain Reaction, Fluorescence, Quantitative RT-PCR

    4) Product Images from "A highly effective and versatile technology for the isolation of RNAs from grapevines and other woody perennials for use in virus diagnostics"

    Article Title: A highly effective and versatile technology for the isolation of RNAs from grapevines and other woody perennials for use in virus diagnostics

    Journal: Virology Journal

    doi: 10.1186/s12985-015-0376-3

    Profile of total RNA isolated by using five commercial kits. a Denaturing gel electrophoresis of total RNA isolated from peach. b Denaturing gel electrophoresis of total RNA from grapevine leaves. 50 mg of young peach and grapevine leaves (indicated as Y), and mature (M) grapevine leaves was used in RNA isolation with Spectrum™ Plant Total RNA kit (Sigma), RNeasy Plant mini kit (Qiagen), Plant/fungi total RNA kit (Norgen), AccuPrep viral RNA extraction kit (Bioneer) and TRIzol Reagent (Life Technologies). The total RNA yield (μg), A260/A280 and A260/A230 ratios averaged from two replicates are given below each gel panel. 28S rRNA, 18S rRNA and small RNAs are indicated with arrows. c Capillary electrophoresis of total RNA with Agilent Bioanalyzer. One ml of each of the total RNA preparations isolated using these five systems was used for the analysis with an Agilent Bioanalyzer 2100 equipped with an RNA Nano chip
    Figure Legend Snippet: Profile of total RNA isolated by using five commercial kits. a Denaturing gel electrophoresis of total RNA isolated from peach. b Denaturing gel electrophoresis of total RNA from grapevine leaves. 50 mg of young peach and grapevine leaves (indicated as Y), and mature (M) grapevine leaves was used in RNA isolation with Spectrum™ Plant Total RNA kit (Sigma), RNeasy Plant mini kit (Qiagen), Plant/fungi total RNA kit (Norgen), AccuPrep viral RNA extraction kit (Bioneer) and TRIzol Reagent (Life Technologies). The total RNA yield (μg), A260/A280 and A260/A230 ratios averaged from two replicates are given below each gel panel. 28S rRNA, 18S rRNA and small RNAs are indicated with arrows. c Capillary electrophoresis of total RNA with Agilent Bioanalyzer. One ml of each of the total RNA preparations isolated using these five systems was used for the analysis with an Agilent Bioanalyzer 2100 equipped with an RNA Nano chip

    Techniques Used: Isolation, Nucleic Acid Electrophoresis, RNA Extraction, Electrophoresis, Chromatin Immunoprecipitation

    5) Product Images from "Differential Roles of Two Homologous Cyclin-Dependent Kinase Inhibitor Genes in Regulating Cell Cycle and Innate Immunity in Arabidopsis 1Differential Roles of Two Homologous Cyclin-Dependent Kinase Inhibitor Genes in Regulating Cell Cycle and Innate Immunity in Arabidopsis 1 [OPEN]"

    Article Title: Differential Roles of Two Homologous Cyclin-Dependent Kinase Inhibitor Genes in Regulating Cell Cycle and Innate Immunity in Arabidopsis 1Differential Roles of Two Homologous Cyclin-Dependent Kinase Inhibitor Genes in Regulating Cell Cycle and Innate Immunity in Arabidopsis 1 [OPEN]

    Journal: Plant Physiology

    doi: 10.1104/pp.15.01466

    smr1-1 accumulation, and cell death in acd6-1 ; 2, acd6-1 ; 3, acd6-1 sim-1 ; 4, acd6-1 smr1-1 ; 5, acd6-1 sim-1 smr1-1 . B, Plant size comparison. Rosette diameters
    Figure Legend Snippet: smr1-1 accumulation, and cell death in acd6-1 ; 2, acd6-1 ; 3, acd6-1 sim-1 ; 4, acd6-1 smr1-1 ; 5, acd6-1 sim-1 smr1-1 . B, Plant size comparison. Rosette diameters

    Techniques Used:

    6) Product Images from "Molecular Cloning and Characterization of Novel Phytocystatin Gene from Turmeric, Curcuma longa"

    Article Title: Molecular Cloning and Characterization of Novel Phytocystatin Gene from Turmeric, Curcuma longa

    Journal: BioMed Research International

    doi: 10.1155/2014/973790

    Total RNA integrity test on 1.0% agarose gel. Intact 28S and 18S total RNA bands can be observed on the agarose gel, indicating a good integrity of the total RNA after being extracted by different methods (lanes 1 and 2: modified CTAB method, lane 3, 4: RNAzol RT, and lane 5, 6: RNeasy Plant Mini kit).
    Figure Legend Snippet: Total RNA integrity test on 1.0% agarose gel. Intact 28S and 18S total RNA bands can be observed on the agarose gel, indicating a good integrity of the total RNA after being extracted by different methods (lanes 1 and 2: modified CTAB method, lane 3, 4: RNAzol RT, and lane 5, 6: RNeasy Plant Mini kit).

    Techniques Used: Agarose Gel Electrophoresis, Modification

    7) Product Images from "A practical examination of RNA isolation methods for European pear (Pyrus communis)"

    Article Title: A practical examination of RNA isolation methods for European pear (Pyrus communis)

    Journal: BMC Research Notes

    doi: 10.1186/s13104-017-2564-2

    Both yield and quality are variable within and across kit based methods, yet the modified CTAB protocol produces consistent high yield and quality in stored ‘d’Anjou tissues. a RINs are higher and more consistent across methods for stored ‘d’Anjou’ peel than cortex. b Excluding protocols with degraded RNA, yields are variable across kits with the highest yield using the CTAB protocol. c Excluding protocols with degraded RNA, A 260/280− ratios were also variable across methods, with CTAB again producing the cleanest RNA. Error bars are standard error of the mean, where applicable. Some data are missing due to very low yield or severely degraded individual samples. QRP RLC Qiagen RNeasy Plant using buffer RLC, CTAB our modified CTAB protocol see Additional file 1 , OHP Omega EZNA HP total RNA, TF thermo fisher, MN RAP Macherey–Nagel NucleoSpin Plant using buffer RAP, OTR Omega EZNA total RNA, QRP RLT Qiagen RNeasy Plant using buffer RLT, MN RA1 Macherey–Nagel NucleoSpin Plant using buffer RA1, ZR ZR plant RNA MiniPrep, OPR Omega EZNA plant RNA Kit 1, QRU Qiagen RNeasy plus universal
    Figure Legend Snippet: Both yield and quality are variable within and across kit based methods, yet the modified CTAB protocol produces consistent high yield and quality in stored ‘d’Anjou tissues. a RINs are higher and more consistent across methods for stored ‘d’Anjou’ peel than cortex. b Excluding protocols with degraded RNA, yields are variable across kits with the highest yield using the CTAB protocol. c Excluding protocols with degraded RNA, A 260/280− ratios were also variable across methods, with CTAB again producing the cleanest RNA. Error bars are standard error of the mean, where applicable. Some data are missing due to very low yield or severely degraded individual samples. QRP RLC Qiagen RNeasy Plant using buffer RLC, CTAB our modified CTAB protocol see Additional file 1 , OHP Omega EZNA HP total RNA, TF thermo fisher, MN RAP Macherey–Nagel NucleoSpin Plant using buffer RAP, OTR Omega EZNA total RNA, QRP RLT Qiagen RNeasy Plant using buffer RLT, MN RA1 Macherey–Nagel NucleoSpin Plant using buffer RA1, ZR ZR plant RNA MiniPrep, OPR Omega EZNA plant RNA Kit 1, QRU Qiagen RNeasy plus universal

    Techniques Used: Modification

    8) Product Images from "Multiplex Detection of Aspergillus fumigatus Mycoviruses"

    Article Title: Multiplex Detection of Aspergillus fumigatus Mycoviruses

    Journal: Viruses

    doi: 10.3390/v10050247

    Conventional PCR to check amplicon size prior to multiplex PCR. Amplicon sizes were checked on two percent agarose gel prior to performing multiplex PCR for A. fumigatus dsRNA mycoviruses. AfuCV, AfuPV-1 and AfuTmV-1 dsRNAs were extracted using LiCl extraction (Lanes 2, 4 and 6, respectively) and RNeasy Plant Mini kit (Lanes 3, 5 and 7, respectively) and used as templates for amplification with AfuCV, AfuPV-1 and AfuTmV-1 primers (2–3, 4–5, 6–7, respectively). Hyperladder-I was used as a marker to estimate the size of the amplicons (Lane 1).
    Figure Legend Snippet: Conventional PCR to check amplicon size prior to multiplex PCR. Amplicon sizes were checked on two percent agarose gel prior to performing multiplex PCR for A. fumigatus dsRNA mycoviruses. AfuCV, AfuPV-1 and AfuTmV-1 dsRNAs were extracted using LiCl extraction (Lanes 2, 4 and 6, respectively) and RNeasy Plant Mini kit (Lanes 3, 5 and 7, respectively) and used as templates for amplification with AfuCV, AfuPV-1 and AfuTmV-1 primers (2–3, 4–5, 6–7, respectively). Hyperladder-I was used as a marker to estimate the size of the amplicons (Lane 1).

    Techniques Used: Polymerase Chain Reaction, Amplification, Multiplex Assay, Agarose Gel Electrophoresis, Marker

    9) Product Images from "Multiplex Detection of Aspergillus fumigatus Mycoviruses"

    Article Title: Multiplex Detection of Aspergillus fumigatus Mycoviruses

    Journal: Viruses

    doi: 10.3390/v10050247

    Conventional PCR to check amplicon size prior to multiplex PCR. Amplicon sizes were checked on two percent agarose gel prior to performing multiplex PCR for A. fumigatus dsRNA mycoviruses. AfuCV, AfuPV-1 and AfuTmV-1 dsRNAs were extracted using LiCl extraction (Lanes 2, 4 and 6, respectively) and RNeasy Plant Mini kit (Lanes 3, 5 and 7, respectively) and used as templates for amplification with AfuCV, AfuPV-1 and AfuTmV-1 primers (2–3, 4–5, 6–7, respectively). Hyperladder-I was used as a marker to estimate the size of the amplicons (Lane 1).
    Figure Legend Snippet: Conventional PCR to check amplicon size prior to multiplex PCR. Amplicon sizes were checked on two percent agarose gel prior to performing multiplex PCR for A. fumigatus dsRNA mycoviruses. AfuCV, AfuPV-1 and AfuTmV-1 dsRNAs were extracted using LiCl extraction (Lanes 2, 4 and 6, respectively) and RNeasy Plant Mini kit (Lanes 3, 5 and 7, respectively) and used as templates for amplification with AfuCV, AfuPV-1 and AfuTmV-1 primers (2–3, 4–5, 6–7, respectively). Hyperladder-I was used as a marker to estimate the size of the amplicons (Lane 1).

    Techniques Used: Polymerase Chain Reaction, Amplification, Multiplex Assay, Agarose Gel Electrophoresis, Marker

    10) Product Images from "A practical examination of RNA isolation methods for European pear (Pyrus communis)"

    Article Title: A practical examination of RNA isolation methods for European pear (Pyrus communis)

    Journal: BMC Research Notes

    doi: 10.1186/s13104-017-2564-2

    Both yield and quality are variable within and across kit based methods, yet the modified CTAB protocol produces consistent high yield and quality in stored ‘d’Anjou tissues. a RINs are higher and more consistent across methods for stored ‘d’Anjou’ peel than cortex. b Excluding protocols with degraded RNA, yields are variable across kits with the highest yield using the CTAB protocol. c Excluding protocols with degraded RNA, A 260/280− ratios were also variable across methods, with CTAB again producing the cleanest RNA. Error bars are standard error of the mean, where applicable. Some data are missing due to very low yield or severely degraded individual samples. QRP RLC Qiagen RNeasy Plant using buffer RLC, CTAB our modified CTAB protocol see Additional file 1 , OHP Omega EZNA HP total RNA, TF thermo fisher, MN RAP Macherey–Nagel NucleoSpin Plant using buffer RAP, OTR Omega EZNA total RNA, QRP RLT Qiagen RNeasy Plant using buffer RLT, MN RA1 Macherey–Nagel NucleoSpin Plant using buffer RA1, ZR ZR plant RNA MiniPrep, OPR Omega EZNA plant RNA Kit 1, QRU Qiagen RNeasy plus universal
    Figure Legend Snippet: Both yield and quality are variable within and across kit based methods, yet the modified CTAB protocol produces consistent high yield and quality in stored ‘d’Anjou tissues. a RINs are higher and more consistent across methods for stored ‘d’Anjou’ peel than cortex. b Excluding protocols with degraded RNA, yields are variable across kits with the highest yield using the CTAB protocol. c Excluding protocols with degraded RNA, A 260/280− ratios were also variable across methods, with CTAB again producing the cleanest RNA. Error bars are standard error of the mean, where applicable. Some data are missing due to very low yield or severely degraded individual samples. QRP RLC Qiagen RNeasy Plant using buffer RLC, CTAB our modified CTAB protocol see Additional file 1 , OHP Omega EZNA HP total RNA, TF thermo fisher, MN RAP Macherey–Nagel NucleoSpin Plant using buffer RAP, OTR Omega EZNA total RNA, QRP RLT Qiagen RNeasy Plant using buffer RLT, MN RA1 Macherey–Nagel NucleoSpin Plant using buffer RA1, ZR ZR plant RNA MiniPrep, OPR Omega EZNA plant RNA Kit 1, QRU Qiagen RNeasy plus universal

    Techniques Used: Modification

    11) Product Images from "A highly effective and versatile technology for the isolation of RNAs from grapevines and other woody perennials for use in virus diagnostics"

    Article Title: A highly effective and versatile technology for the isolation of RNAs from grapevines and other woody perennials for use in virus diagnostics

    Journal: Virology Journal

    doi: 10.1186/s12985-015-0376-3

    Profile of total RNA isolated by using five commercial kits. a Denaturing gel electrophoresis of total RNA isolated from peach. b Denaturing gel electrophoresis of total RNA from grapevine leaves. 50 mg of young peach and grapevine leaves (indicated as Y), and mature (M) grapevine leaves was used in RNA isolation with Spectrum™ Plant Total RNA kit (Sigma), RNeasy Plant mini kit (Qiagen), Plant/fungi total RNA kit (Norgen), AccuPrep viral RNA extraction kit (Bioneer) and TRIzol Reagent (Life Technologies). The total RNA yield (μg), A260/A280 and A260/A230 ratios averaged from two replicates are given below each gel panel. 28S rRNA, 18S rRNA and small RNAs are indicated with arrows. c Capillary electrophoresis of total RNA with Agilent Bioanalyzer. One ml of each of the total RNA preparations isolated using these five systems was used for the analysis with an Agilent Bioanalyzer 2100 equipped with an RNA Nano chip
    Figure Legend Snippet: Profile of total RNA isolated by using five commercial kits. a Denaturing gel electrophoresis of total RNA isolated from peach. b Denaturing gel electrophoresis of total RNA from grapevine leaves. 50 mg of young peach and grapevine leaves (indicated as Y), and mature (M) grapevine leaves was used in RNA isolation with Spectrum™ Plant Total RNA kit (Sigma), RNeasy Plant mini kit (Qiagen), Plant/fungi total RNA kit (Norgen), AccuPrep viral RNA extraction kit (Bioneer) and TRIzol Reagent (Life Technologies). The total RNA yield (μg), A260/A280 and A260/A230 ratios averaged from two replicates are given below each gel panel. 28S rRNA, 18S rRNA and small RNAs are indicated with arrows. c Capillary electrophoresis of total RNA with Agilent Bioanalyzer. One ml of each of the total RNA preparations isolated using these five systems was used for the analysis with an Agilent Bioanalyzer 2100 equipped with an RNA Nano chip

    Techniques Used: Isolation, Nucleic Acid Electrophoresis, RNA Extraction, Electrophoresis, Chromatin Immunoprecipitation

    12) Product Images from "Phragmoplast Orienting Kinesin 2 Is a Weak Motor Switching between Processive and Diffusive Modes"

    Article Title: Phragmoplast Orienting Kinesin 2 Is a Weak Motor Switching between Processive and Diffusive Modes

    Journal: Biophysical Journal

    doi: 10.1016/j.bpj.2018.06.012

    Directionality of the phragmoplast peripheral microtubules. Arabidopsis root meristem stably expressing 35S:GFP-MBD and pUBQ:RFP-EB1b during late cytokinesis is shown. Plus ends (RFP-EB1b) marked with arrow heads of peripheral microtubules (GFP-MBD) from
    Figure Legend Snippet: Directionality of the phragmoplast peripheral microtubules. Arabidopsis root meristem stably expressing 35S:GFP-MBD and pUBQ:RFP-EB1b during late cytokinesis is shown. Plus ends (RFP-EB1b) marked with arrow heads of peripheral microtubules (GFP-MBD) from

    Techniques Used: Stable Transfection, Expressing

    13) Product Images from "A practical examination of RNA isolation methods for European pear (Pyrus communis)"

    Article Title: A practical examination of RNA isolation methods for European pear (Pyrus communis)

    Journal: BMC Research Notes

    doi: 10.1186/s13104-017-2564-2

    Both yield and quality are variable within and across kit based methods, yet the modified CTAB protocol produces consistent high yield and quality in stored ‘d’Anjou tissues. a RINs are higher and more consistent across methods for stored ‘d’Anjou’ peel than cortex. b Excluding protocols with degraded RNA, yields are variable across kits with the highest yield using the CTAB protocol. c Excluding protocols with degraded RNA, A 260/280− ratios were also variable across methods, with CTAB again producing the cleanest RNA. Error bars are standard error of the mean, where applicable. Some data are missing due to very low yield or severely degraded individual samples. QRP RLC Qiagen RNeasy Plant using buffer RLC, CTAB our modified CTAB protocol see Additional file 1 , OHP Omega EZNA HP total RNA, TF thermo fisher, MN RAP Macherey–Nagel NucleoSpin Plant using buffer RAP, OTR Omega EZNA total RNA, QRP RLT Qiagen RNeasy Plant using buffer RLT, MN RA1 Macherey–Nagel NucleoSpin Plant using buffer RA1, ZR ZR plant RNA MiniPrep, OPR Omega EZNA plant RNA Kit 1, QRU Qiagen RNeasy plus universal
    Figure Legend Snippet: Both yield and quality are variable within and across kit based methods, yet the modified CTAB protocol produces consistent high yield and quality in stored ‘d’Anjou tissues. a RINs are higher and more consistent across methods for stored ‘d’Anjou’ peel than cortex. b Excluding protocols with degraded RNA, yields are variable across kits with the highest yield using the CTAB protocol. c Excluding protocols with degraded RNA, A 260/280− ratios were also variable across methods, with CTAB again producing the cleanest RNA. Error bars are standard error of the mean, where applicable. Some data are missing due to very low yield or severely degraded individual samples. QRP RLC Qiagen RNeasy Plant using buffer RLC, CTAB our modified CTAB protocol see Additional file 1 , OHP Omega EZNA HP total RNA, TF thermo fisher, MN RAP Macherey–Nagel NucleoSpin Plant using buffer RAP, OTR Omega EZNA total RNA, QRP RLT Qiagen RNeasy Plant using buffer RLT, MN RA1 Macherey–Nagel NucleoSpin Plant using buffer RA1, ZR ZR plant RNA MiniPrep, OPR Omega EZNA plant RNA Kit 1, QRU Qiagen RNeasy plus universal

    Techniques Used: Modification

    14) Product Images from "Techniques for the Isolation of High-Quality RNA from Cells Encapsulated in Chitosan Hydrogels"

    Article Title: Techniques for the Isolation of High-Quality RNA from Cells Encapsulated in Chitosan Hydrogels

    Journal: Tissue Engineering. Part C, Methods

    doi: 10.1089/ten.tec.2012.0693

    Representative end-point RT-PCR gene expression results for (a) 18S and (b) TfR from RNA isolated using the freeze grind+CTAB+RNeasy ® (FCR) method, the mince+CTAB+RNeasy ® (MCR) method, and the lysozyme+CTAB+RNeasy ® (LCR) method. Genomic contamination was detected following agarose gel electrophoresis in all minus-RT controls. In addition, as shown in the TfR results, where the primers were designed to span an intron–exon boundary, two products were formed during the PCR, corresponding to a genomic product size of 270 bp and an mRNA product size of 62 bp.
    Figure Legend Snippet: Representative end-point RT-PCR gene expression results for (a) 18S and (b) TfR from RNA isolated using the freeze grind+CTAB+RNeasy ® (FCR) method, the mince+CTAB+RNeasy ® (MCR) method, and the lysozyme+CTAB+RNeasy ® (LCR) method. Genomic contamination was detected following agarose gel electrophoresis in all minus-RT controls. In addition, as shown in the TfR results, where the primers were designed to span an intron–exon boundary, two products were formed during the PCR, corresponding to a genomic product size of 270 bp and an mRNA product size of 62 bp.

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Expressing, Isolation, Agarose Gel Electrophoresis, Polymerase Chain Reaction

    15) Product Images from "A Key ABA Catabolic Gene, OsABA8ox3, Is Involved in Drought Stress Resistance in Rice"

    Article Title: A Key ABA Catabolic Gene, OsABA8ox3, Is Involved in Drought Stress Resistance in Rice

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0116646

    Relative expression levels of 9 stress-related genes in the WT and OsABA8ox3 RNAi transgenic plants under normal and PEG-treated conditions detected by qRT-PCR. Four-leaf stage seedlings were treated with 20% PEG for 2 h, and then the samples were quickly frozen with liquid nitrogen for RNA extraction. The genes include dehydration-responsive element-binding protein (Os06g03670), zinc finger protein (Os05g10670), late embryogenesis abundant proteins (Os05g46480, Os01g50910 and Os04g49980), dehydrin family proteins (Os11g26760 and Os01g50700) and heat shock protein (Os03g16920 and Os02g54140).
    Figure Legend Snippet: Relative expression levels of 9 stress-related genes in the WT and OsABA8ox3 RNAi transgenic plants under normal and PEG-treated conditions detected by qRT-PCR. Four-leaf stage seedlings were treated with 20% PEG for 2 h, and then the samples were quickly frozen with liquid nitrogen for RNA extraction. The genes include dehydration-responsive element-binding protein (Os06g03670), zinc finger protein (Os05g10670), late embryogenesis abundant proteins (Os05g46480, Os01g50910 and Os04g49980), dehydrin family proteins (Os11g26760 and Os01g50700) and heat shock protein (Os03g16920 and Os02g54140).

    Techniques Used: Expressing, Transgenic Assay, Quantitative RT-PCR, RNA Extraction, Binding Assay

    16) Product Images from "Subcellular Localization and Rolling Circle Replication of Peach Latent Mosaic Viroid: Hallmarks of Group A Viroids"

    Article Title: Subcellular Localization and Rolling Circle Replication of Peach Latent Mosaic Viroid: Hallmarks of Group A Viroids

    Journal: Journal of Virology

    doi:

    Autoradiograms of Northern blot hybridizations of RNA samples isolated from both healthy and PLMVd-infected peach leaves. RNA samples were fractionated on either agarose (A to C) or polyacrylamide (D and E) gels and then blotted onto nylon filters. The polarity of the PLMVd riboprobe is indicated by the symbol (+) and (−) at the top of the panel. (A) RNA samples were isolated by various extraction procedures: RNeasy Plant mini kit (Qiagen) in the presence of EDTA (lanes 3 to 5); Tris-EDTA isolation (lanes 6 and 7); RNeasy Plant mini kit in the absence of additional EDTA (lane 8); and the PEG precipitation procedure (lanes 9 to 12). In lanes 1 and 2, nonradioactive synthetic PLMVd transcripts of plus (761 and 588 nt) and minus (745 and 462 nt) polarity, respectively, were loaded as controls. Lane 3 contains a sample of healthy GF-305 peach cultivar; lanes 4, 6, 8, 9, 11, and 12 contain RNA samples isolated from a PLMVd-infected RedGold peach cultivar. The samples in lanes 11 and 12 were subjected to either DNase treatment or alkaline hydrolysis prior electrophoresis. Lanes 5 and 7 contain to samples from a PLMVd-infected Redhaven cultivar, while lane 10 contains a sample from the leaves of the Agua cultivar. (B and C) The same filter following hybridization with either the plus- or minus-polarity riboprobe. All samples were isolated by the PEG precipitation procedure. Lanes 1 and 2 contain samples isolated from different leaves of a Redhaven peach. Lane 3 contains an RNA sample from a healthy Bailey cultivar. Lanes 4 and 5 contain RNA samples of PLMVd-infected Redhaven and Siberian C cultivars. (D and E) Filter following PAGE and blotting that was probed with both the plus- and minus-polarity riboprobes, respectively. In panel D, lanes 1 and 2 contain synthetic circular and linear PLMVd controls, while lanes 3 to 5 (which correspond to lanes 1 to 3 of panel E) contain samples from PLMVd-infected Redgold, Hardired, and Siberian C cultivars and lanes 6 and 7 (which correspond to lanes 4 and 5 of panel E) contain samples from healthy GF-305 and Bailey cultivars. Adjacent to the gel, the positions of several PLMVd transcripts are indicated as size references including both the circular (338C) and linear (338L) strands and the mixture of circular and linear molecules (PLMVd). The position of the unknown ∼1-kb species is also indicated in panels A and B.
    Figure Legend Snippet: Autoradiograms of Northern blot hybridizations of RNA samples isolated from both healthy and PLMVd-infected peach leaves. RNA samples were fractionated on either agarose (A to C) or polyacrylamide (D and E) gels and then blotted onto nylon filters. The polarity of the PLMVd riboprobe is indicated by the symbol (+) and (−) at the top of the panel. (A) RNA samples were isolated by various extraction procedures: RNeasy Plant mini kit (Qiagen) in the presence of EDTA (lanes 3 to 5); Tris-EDTA isolation (lanes 6 and 7); RNeasy Plant mini kit in the absence of additional EDTA (lane 8); and the PEG precipitation procedure (lanes 9 to 12). In lanes 1 and 2, nonradioactive synthetic PLMVd transcripts of plus (761 and 588 nt) and minus (745 and 462 nt) polarity, respectively, were loaded as controls. Lane 3 contains a sample of healthy GF-305 peach cultivar; lanes 4, 6, 8, 9, 11, and 12 contain RNA samples isolated from a PLMVd-infected RedGold peach cultivar. The samples in lanes 11 and 12 were subjected to either DNase treatment or alkaline hydrolysis prior electrophoresis. Lanes 5 and 7 contain to samples from a PLMVd-infected Redhaven cultivar, while lane 10 contains a sample from the leaves of the Agua cultivar. (B and C) The same filter following hybridization with either the plus- or minus-polarity riboprobe. All samples were isolated by the PEG precipitation procedure. Lanes 1 and 2 contain samples isolated from different leaves of a Redhaven peach. Lane 3 contains an RNA sample from a healthy Bailey cultivar. Lanes 4 and 5 contain RNA samples of PLMVd-infected Redhaven and Siberian C cultivars. (D and E) Filter following PAGE and blotting that was probed with both the plus- and minus-polarity riboprobes, respectively. In panel D, lanes 1 and 2 contain synthetic circular and linear PLMVd controls, while lanes 3 to 5 (which correspond to lanes 1 to 3 of panel E) contain samples from PLMVd-infected Redgold, Hardired, and Siberian C cultivars and lanes 6 and 7 (which correspond to lanes 4 and 5 of panel E) contain samples from healthy GF-305 and Bailey cultivars. Adjacent to the gel, the positions of several PLMVd transcripts are indicated as size references including both the circular (338C) and linear (338L) strands and the mixture of circular and linear molecules (PLMVd). The position of the unknown ∼1-kb species is also indicated in panels A and B.

    Techniques Used: Northern Blot, Isolation, Infection, Electrophoresis, Hybridization, Polyacrylamide Gel Electrophoresis

    17) Product Images from "A method for extracting high-quality total RNA from plant rich in polysaccharides and polyphenols using Dendrobium huoshanense"

    Article Title: A method for extracting high-quality total RNA from plant rich in polysaccharides and polyphenols using Dendrobium huoshanense

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0196592

    1.0% agarose gel electrophoresis of total RNA isolated. A, three intact RNA bands for 28S, 18S and 5S RNA. Lane 1, lane 2 and lane 3 in A, B, C, D and E contain 1 μg of total RNA from D . huoshanense stem, leaf and flower, respectively. A: modified CHAN method; B: original CHAN method; C: Trizol method; D: RNeasy Plant Mini Kit method; E: RNAprep Pure Plant Kit method.
    Figure Legend Snippet: 1.0% agarose gel electrophoresis of total RNA isolated. A, three intact RNA bands for 28S, 18S and 5S RNA. Lane 1, lane 2 and lane 3 in A, B, C, D and E contain 1 μg of total RNA from D . huoshanense stem, leaf and flower, respectively. A: modified CHAN method; B: original CHAN method; C: Trizol method; D: RNeasy Plant Mini Kit method; E: RNAprep Pure Plant Kit method.

    Techniques Used: Agarose Gel Electrophoresis, Isolation, Modification

    18) Product Images from "Efficient and High-Quality RNA Isolation from Metabolite-Rich Tissues of Stevia rebaudiana, an Important Commercial Crop"

    Article Title: Efficient and High-Quality RNA Isolation from Metabolite-Rich Tissues of Stevia rebaudiana, an Important Commercial Crop

    Journal: Tropical Life Sciences Research

    doi: 10.21315/tlsr2019.30.1.9

    Agarose gel electrophoresis and the respective electropherograms of the total RNA samples extracted from (a) leaves, and (b) stems, of Stevia rebaudiana . Two distinct rRNA bands and peaks were clearly observed, for each sample, in both the gel and electropherograms respectively, signifying high RNA quality and minimal degradation. LT1-LT3: Replicates of RNA extracts from leaf tissues (400–600 ng); ST1-ST3: Replicates of RNA extracts from stem tissues (300–400 ng).
    Figure Legend Snippet: Agarose gel electrophoresis and the respective electropherograms of the total RNA samples extracted from (a) leaves, and (b) stems, of Stevia rebaudiana . Two distinct rRNA bands and peaks were clearly observed, for each sample, in both the gel and electropherograms respectively, signifying high RNA quality and minimal degradation. LT1-LT3: Replicates of RNA extracts from leaf tissues (400–600 ng); ST1-ST3: Replicates of RNA extracts from stem tissues (300–400 ng).

    Techniques Used: Agarose Gel Electrophoresis

    19) Product Images from "Molecular Cloning and Characterization of Novel Phytocystatin Gene from Turmeric, Curcuma longa"

    Article Title: Molecular Cloning and Characterization of Novel Phytocystatin Gene from Turmeric, Curcuma longa

    Journal: BioMed Research International

    doi: 10.1155/2014/973790

    Total RNA integrity test on 1.0% agarose gel. Intact 28S and 18S total RNA bands can be observed on the agarose gel, indicating a good integrity of the total RNA after being extracted by different methods (lanes 1 and 2: modified CTAB method, lane 3, 4: RNAzol RT, and lane 5, 6: RNeasy Plant Mini kit).
    Figure Legend Snippet: Total RNA integrity test on 1.0% agarose gel. Intact 28S and 18S total RNA bands can be observed on the agarose gel, indicating a good integrity of the total RNA after being extracted by different methods (lanes 1 and 2: modified CTAB method, lane 3, 4: RNAzol RT, and lane 5, 6: RNeasy Plant Mini kit).

    Techniques Used: Agarose Gel Electrophoresis, Modification

    20) Product Images from "Molecular Cloning and Characterization of Novel Phytocystatin Gene from Turmeric, Curcuma longa"

    Article Title: Molecular Cloning and Characterization of Novel Phytocystatin Gene from Turmeric, Curcuma longa

    Journal: BioMed Research International

    doi: 10.1155/2014/973790

    Total RNA integrity test on 1.0% agarose gel. Intact 28S and 18S total RNA bands can be observed on the agarose gel, indicating a good integrity of the total RNA after being extracted by different methods (lanes 1 and 2: modified CTAB method, lane 3, 4: RNAzol RT, and lane 5, 6: RNeasy Plant Mini kit).
    Figure Legend Snippet: Total RNA integrity test on 1.0% agarose gel. Intact 28S and 18S total RNA bands can be observed on the agarose gel, indicating a good integrity of the total RNA after being extracted by different methods (lanes 1 and 2: modified CTAB method, lane 3, 4: RNAzol RT, and lane 5, 6: RNeasy Plant Mini kit).

    Techniques Used: Agarose Gel Electrophoresis, Modification

    21) Product Images from "Filter paper-based spin column method for cost-efficient DNA or RNA purification"

    Article Title: Filter paper-based spin column method for cost-efficient DNA or RNA purification

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0203011

    The efficiency of filter paper for purification of nucleic acids from various sources using respective Qiagen kits. (A) Tomato genomic DNAs purified using Qiagen DNeasy plant mini kit. (B) Tomato total RNAs purified using Qiagen RNeasy plant mini kit. (C) PCR products of a GUS fragment purified using Qiagen QIAquick PCR purification kit. (D) PCR products of GUS fragment recovered from an agarose gel using a Qiagen QIAquick gel extraction kit. (E) pUC -19 plasmid DNAs purified using a Qiagen QIAprep spin miniprep kit. For each panel, from left to right are (Q) nucleic acid purified in experiments using original Qiagen spin column, (G) reassembled spin column using two layers of Whatman glass microfiber filters (Grade GF/F), and (P) reassembled spin column using two layers of Whatman qualitative filter paper, (Grade 3) respectively. Upper panel is quantification data based on three experimental replicates normalized according to performance of the Qiagen kit; lower panel is an image of agarose gel electrophoresis for the same volume of purified nucleic acids.
    Figure Legend Snippet: The efficiency of filter paper for purification of nucleic acids from various sources using respective Qiagen kits. (A) Tomato genomic DNAs purified using Qiagen DNeasy plant mini kit. (B) Tomato total RNAs purified using Qiagen RNeasy plant mini kit. (C) PCR products of a GUS fragment purified using Qiagen QIAquick PCR purification kit. (D) PCR products of GUS fragment recovered from an agarose gel using a Qiagen QIAquick gel extraction kit. (E) pUC -19 plasmid DNAs purified using a Qiagen QIAprep spin miniprep kit. For each panel, from left to right are (Q) nucleic acid purified in experiments using original Qiagen spin column, (G) reassembled spin column using two layers of Whatman glass microfiber filters (Grade GF/F), and (P) reassembled spin column using two layers of Whatman qualitative filter paper, (Grade 3) respectively. Upper panel is quantification data based on three experimental replicates normalized according to performance of the Qiagen kit; lower panel is an image of agarose gel electrophoresis for the same volume of purified nucleic acids.

    Techniques Used: Purification, Polymerase Chain Reaction, Agarose Gel Electrophoresis, Gel Extraction, Plasmid Preparation

    Evaluation of purification of tobacco genomic DNA and total RNA using filter paper-based spin columns with respective Qiagen kit buffers and homemade buffers. (A) Agarose gel electrophoresis for 2.5 μl tobacco genomic DNAs elution from purification experiments using Qiagen DNeasy plant mini kit buffers with Qiagen original spin column (Lane Q/Q), filter paper recharged used spin column (Lane Q/R) and filter paper-based homemade spin column (Lane Q/H*), followed by tobacco genomic DNAs purified using homemade buffer with Qiagen original spin column (Lane H/Q), filter paper recharged used spin column (Lane H/R) and filter paper-based homemade spin column (Lane H/H*). (B) UV spectrum curve of tobacco DNAs purified using Qiagen kit (Q/Q, black curve), filter paper recharged spin columns with Qiagen kit buffers (Q/R, blue curve) or homemade buffers (H/R, red curve) from the same amount leaf tissue. Y-axis is UV absorbance, and X-axis is wavelength (nM). (C) Amplification plots for three duplicated qPCR reactions contain 20 ng DNA purified using Qiagen kit (Q/Q, Blue curves) or DNA purified from filter paper recharged spin column with homemade buffer (H/R, Red curves) respectively. The x-axis is PCR cycle numbers, Y-axis is the level of SYBR fluorescence, and the green line is an arbitrary threshold to determine the Cq value (the fractional cycle number at which amplification curve meet threshold level). (D) MOPS-formaldehyde denaturing agarose gel electrophoresis separated 5 μl RNA purified using Qiagen RNeasy plant mini kit buffers with a Qiagen original spin column (Lane Q/Q), filter paper recharged used spin column (Lane Q/R) and homemade filter paper-based spin column (Lane Q/H*), followed total tobacco RNAs purified by using homemade buffer with Qiagen original spin column (Lane H/Q), filter paper recharged used spin column (Lane H/R) and filter paper-based homemade spin column (Lane H/H*). (E) UV spectrum of tobacco total RNA purified using Qiagen kit (Q/Q, black curve), filter paper recharged spin column with Qiagen RNeasy plant mini kit buffers (Q/R, blue curve) or homemade buffers (H/R, red curve). Y-axis is UV absorbance, and the X-axis is wavelength. (F) Amplification plots of three duplicated qRT-PCR reactions for 2.5 ng RNA purified using Qiagen kit (Q/Q, Blue curves) or RNA purified using filter paper recharged spin column with homemade buffer (H/R, Red curves) respectively. Note: * The starting material amount is 100 mg tobacco leaf tissue for experiments using a Qiagen spin column or filter paper recharged spin column, and half amount of plant sample (50 mg) used for homemade spin column purification. All DNAs or RNAs were eluted using 100 ul elution solution.
    Figure Legend Snippet: Evaluation of purification of tobacco genomic DNA and total RNA using filter paper-based spin columns with respective Qiagen kit buffers and homemade buffers. (A) Agarose gel electrophoresis for 2.5 μl tobacco genomic DNAs elution from purification experiments using Qiagen DNeasy plant mini kit buffers with Qiagen original spin column (Lane Q/Q), filter paper recharged used spin column (Lane Q/R) and filter paper-based homemade spin column (Lane Q/H*), followed by tobacco genomic DNAs purified using homemade buffer with Qiagen original spin column (Lane H/Q), filter paper recharged used spin column (Lane H/R) and filter paper-based homemade spin column (Lane H/H*). (B) UV spectrum curve of tobacco DNAs purified using Qiagen kit (Q/Q, black curve), filter paper recharged spin columns with Qiagen kit buffers (Q/R, blue curve) or homemade buffers (H/R, red curve) from the same amount leaf tissue. Y-axis is UV absorbance, and X-axis is wavelength (nM). (C) Amplification plots for three duplicated qPCR reactions contain 20 ng DNA purified using Qiagen kit (Q/Q, Blue curves) or DNA purified from filter paper recharged spin column with homemade buffer (H/R, Red curves) respectively. The x-axis is PCR cycle numbers, Y-axis is the level of SYBR fluorescence, and the green line is an arbitrary threshold to determine the Cq value (the fractional cycle number at which amplification curve meet threshold level). (D) MOPS-formaldehyde denaturing agarose gel electrophoresis separated 5 μl RNA purified using Qiagen RNeasy plant mini kit buffers with a Qiagen original spin column (Lane Q/Q), filter paper recharged used spin column (Lane Q/R) and homemade filter paper-based spin column (Lane Q/H*), followed total tobacco RNAs purified by using homemade buffer with Qiagen original spin column (Lane H/Q), filter paper recharged used spin column (Lane H/R) and filter paper-based homemade spin column (Lane H/H*). (E) UV spectrum of tobacco total RNA purified using Qiagen kit (Q/Q, black curve), filter paper recharged spin column with Qiagen RNeasy plant mini kit buffers (Q/R, blue curve) or homemade buffers (H/R, red curve). Y-axis is UV absorbance, and the X-axis is wavelength. (F) Amplification plots of three duplicated qRT-PCR reactions for 2.5 ng RNA purified using Qiagen kit (Q/Q, Blue curves) or RNA purified using filter paper recharged spin column with homemade buffer (H/R, Red curves) respectively. Note: * The starting material amount is 100 mg tobacco leaf tissue for experiments using a Qiagen spin column or filter paper recharged spin column, and half amount of plant sample (50 mg) used for homemade spin column purification. All DNAs or RNAs were eluted using 100 ul elution solution.

    Techniques Used: Purification, Agarose Gel Electrophoresis, Amplification, Real-time Polymerase Chain Reaction, Polymerase Chain Reaction, Fluorescence, Quantitative RT-PCR

    22) Product Images from "Filter paper-based spin column method for cost-efficient DNA or RNA purification"

    Article Title: Filter paper-based spin column method for cost-efficient DNA or RNA purification

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0203011

    The efficiency of filter paper for purification of nucleic acids from various sources using respective Qiagen kits. (A) Tomato genomic DNAs purified using Qiagen DNeasy plant mini kit. (B) Tomato total RNAs purified using Qiagen RNeasy plant mini kit. (C) PCR products of a GUS fragment purified using Qiagen QIAquick PCR purification kit. (D) PCR products of GUS fragment recovered from an agarose gel using a Qiagen QIAquick gel extraction kit. (E) pUC -19 plasmid DNAs purified using a Qiagen QIAprep spin miniprep kit. For each panel, from left to right are (Q) nucleic acid purified in experiments using original Qiagen spin column, (G) reassembled spin column using two layers of Whatman glass microfiber filters (Grade GF/F), and (P) reassembled spin column using two layers of Whatman qualitative filter paper, (Grade 3) respectively. Upper panel is quantification data based on three experimental replicates normalized according to performance of the Qiagen kit; lower panel is an image of agarose gel electrophoresis for the same volume of purified nucleic acids.
    Figure Legend Snippet: The efficiency of filter paper for purification of nucleic acids from various sources using respective Qiagen kits. (A) Tomato genomic DNAs purified using Qiagen DNeasy plant mini kit. (B) Tomato total RNAs purified using Qiagen RNeasy plant mini kit. (C) PCR products of a GUS fragment purified using Qiagen QIAquick PCR purification kit. (D) PCR products of GUS fragment recovered from an agarose gel using a Qiagen QIAquick gel extraction kit. (E) pUC -19 plasmid DNAs purified using a Qiagen QIAprep spin miniprep kit. For each panel, from left to right are (Q) nucleic acid purified in experiments using original Qiagen spin column, (G) reassembled spin column using two layers of Whatman glass microfiber filters (Grade GF/F), and (P) reassembled spin column using two layers of Whatman qualitative filter paper, (Grade 3) respectively. Upper panel is quantification data based on three experimental replicates normalized according to performance of the Qiagen kit; lower panel is an image of agarose gel electrophoresis for the same volume of purified nucleic acids.

    Techniques Used: Purification, Polymerase Chain Reaction, Agarose Gel Electrophoresis, Gel Extraction, Plasmid Preparation

    23) Product Images from "LogSpin: a simple, economical and fast method for RNA isolation from infected or healthy plants and other eukaryotic tissues"

    Article Title: LogSpin: a simple, economical and fast method for RNA isolation from infected or healthy plants and other eukaryotic tissues

    Journal: BMC Research Notes

    doi: 10.1186/1756-0500-5-45

    RNA separation on 1% agarose gel . A : RNA extracted from Arabidopsis leaves. Lane 1, RNA extracted using Qiagen RNeasy kit (kit) and lane 2, RNA extracted by LogSpin protocol. B : RNA extracted from B. cinerea (Botrytis) or A. brassicicola (Alternaria) mycelium. Lanes 1 and 3, RNA extracted using Norgen Plant/Fungi RNA Purification kit (kit). Lanes 2 and 4, RNA extracted using LogSpin protocol. C : Qualitative assessment of the integrity of a total RNA sample extracted using LogSpin protocol from tomato leaves by bioanalyzer. M (bp), DNA ladder in base pairs; kit, gel provided by the bioanalyzer kit.
    Figure Legend Snippet: RNA separation on 1% agarose gel . A : RNA extracted from Arabidopsis leaves. Lane 1, RNA extracted using Qiagen RNeasy kit (kit) and lane 2, RNA extracted by LogSpin protocol. B : RNA extracted from B. cinerea (Botrytis) or A. brassicicola (Alternaria) mycelium. Lanes 1 and 3, RNA extracted using Norgen Plant/Fungi RNA Purification kit (kit). Lanes 2 and 4, RNA extracted using LogSpin protocol. C : Qualitative assessment of the integrity of a total RNA sample extracted using LogSpin protocol from tomato leaves by bioanalyzer. M (bp), DNA ladder in base pairs; kit, gel provided by the bioanalyzer kit.

    Techniques Used: Agarose Gel Electrophoresis, Purification

    24) Product Images from "Molecular Cloning and Characterization of Novel Phytocystatin Gene from Turmeric, Curcuma longa"

    Article Title: Molecular Cloning and Characterization of Novel Phytocystatin Gene from Turmeric, Curcuma longa

    Journal: BioMed Research International

    doi: 10.1155/2014/973790

    Total RNA integrity test on 1.0% agarose gel. Intact 28S and 18S total RNA bands can be observed on the agarose gel, indicating a good integrity of the total RNA after being extracted by different methods (lanes 1 and 2: modified CTAB method, lane 3, 4: RNAzol RT, and lane 5, 6: RNeasy Plant Mini kit).
    Figure Legend Snippet: Total RNA integrity test on 1.0% agarose gel. Intact 28S and 18S total RNA bands can be observed on the agarose gel, indicating a good integrity of the total RNA after being extracted by different methods (lanes 1 and 2: modified CTAB method, lane 3, 4: RNAzol RT, and lane 5, 6: RNeasy Plant Mini kit).

    Techniques Used: Agarose Gel Electrophoresis, Modification

    25) Product Images from "One-Carbon Metabolism in Plants. Regulation of Tetrahydrofolate Synthesis during Germination and Seedling Development"

    Article Title: One-Carbon Metabolism in Plants. Regulation of Tetrahydrofolate Synthesis during Germination and Seedling Development

    Journal: Plant Physiology

    doi: 10.1104/pp.016915

    Quantification of the HPPK-DHPS mRNA by semiquantitative RT-PCR. Total RNA was isolated from 6-d-old green pea leaves using the RNeasy Plant Mini Kit (Qiagen USA, Valencia, CA), reverse transcribed, and PCR-amplified with primers specific for HPPK-DHPS. Templates amplified for 20 to 28 cycles were analyzed by Southern blot, and signal intensity was quantified using a phosphor imager (background values are around 10 3 counts mm −2 ). The insert corresponds to the scanned image obtained in the log-linear range of amplification of the target gene.
    Figure Legend Snippet: Quantification of the HPPK-DHPS mRNA by semiquantitative RT-PCR. Total RNA was isolated from 6-d-old green pea leaves using the RNeasy Plant Mini Kit (Qiagen USA, Valencia, CA), reverse transcribed, and PCR-amplified with primers specific for HPPK-DHPS. Templates amplified for 20 to 28 cycles were analyzed by Southern blot, and signal intensity was quantified using a phosphor imager (background values are around 10 3 counts mm −2 ). The insert corresponds to the scanned image obtained in the log-linear range of amplification of the target gene.

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Isolation, Polymerase Chain Reaction, Amplification, Southern Blot

    26) Product Images from "Multiplex Detection of Aspergillus fumigatus Mycoviruses"

    Article Title: Multiplex Detection of Aspergillus fumigatus Mycoviruses

    Journal: Viruses

    doi: 10.3390/v10050247

    Conventional PCR to check amplicon size prior to multiplex PCR. Amplicon sizes were checked on two percent agarose gel prior to performing multiplex PCR for A. fumigatus dsRNA mycoviruses. AfuCV, AfuPV-1 and AfuTmV-1 dsRNAs were extracted using LiCl extraction (Lanes 2, 4 and 6, respectively) and RNeasy Plant Mini kit (Lanes 3, 5 and 7, respectively) and used as templates for amplification with AfuCV, AfuPV-1 and AfuTmV-1 primers (2–3, 4–5, 6–7, respectively). Hyperladder-I was used as a marker to estimate the size of the amplicons (Lane 1).
    Figure Legend Snippet: Conventional PCR to check amplicon size prior to multiplex PCR. Amplicon sizes were checked on two percent agarose gel prior to performing multiplex PCR for A. fumigatus dsRNA mycoviruses. AfuCV, AfuPV-1 and AfuTmV-1 dsRNAs were extracted using LiCl extraction (Lanes 2, 4 and 6, respectively) and RNeasy Plant Mini kit (Lanes 3, 5 and 7, respectively) and used as templates for amplification with AfuCV, AfuPV-1 and AfuTmV-1 primers (2–3, 4–5, 6–7, respectively). Hyperladder-I was used as a marker to estimate the size of the amplicons (Lane 1).

    Techniques Used: Polymerase Chain Reaction, Amplification, Multiplex Assay, Agarose Gel Electrophoresis, Marker

    27) Product Images from "A practical examination of RNA isolation methods for European pear (Pyrus communis)"

    Article Title: A practical examination of RNA isolation methods for European pear (Pyrus communis)

    Journal: BMC Research Notes

    doi: 10.1186/s13104-017-2564-2

    Both yield and quality are variable within and across kit based methods, yet the modified CTAB protocol produces consistent high yield and quality in stored ‘d’Anjou tissues. a RINs are higher and more consistent across methods for stored ‘d’Anjou’ peel than cortex. b Excluding protocols with degraded RNA, yields are variable across kits with the highest yield using the CTAB protocol. c Excluding protocols with degraded RNA, A 260/280− ratios were also variable across methods, with CTAB again producing the cleanest RNA. Error bars are standard error of the mean, where applicable. Some data are missing due to very low yield or severely degraded individual samples. QRP RLC Qiagen RNeasy Plant using buffer RLC, CTAB our modified CTAB protocol see Additional file 1 , OHP Omega EZNA HP total RNA, TF thermo fisher, MN RAP Macherey–Nagel NucleoSpin Plant using buffer RAP, OTR Omega EZNA total RNA, QRP RLT Qiagen RNeasy Plant using buffer RLT, MN RA1 Macherey–Nagel NucleoSpin Plant using buffer RA1, ZR ZR plant RNA MiniPrep, OPR Omega EZNA plant RNA Kit 1, QRU Qiagen RNeasy plus universal
    Figure Legend Snippet: Both yield and quality are variable within and across kit based methods, yet the modified CTAB protocol produces consistent high yield and quality in stored ‘d’Anjou tissues. a RINs are higher and more consistent across methods for stored ‘d’Anjou’ peel than cortex. b Excluding protocols with degraded RNA, yields are variable across kits with the highest yield using the CTAB protocol. c Excluding protocols with degraded RNA, A 260/280− ratios were also variable across methods, with CTAB again producing the cleanest RNA. Error bars are standard error of the mean, where applicable. Some data are missing due to very low yield or severely degraded individual samples. QRP RLC Qiagen RNeasy Plant using buffer RLC, CTAB our modified CTAB protocol see Additional file 1 , OHP Omega EZNA HP total RNA, TF thermo fisher, MN RAP Macherey–Nagel NucleoSpin Plant using buffer RAP, OTR Omega EZNA total RNA, QRP RLT Qiagen RNeasy Plant using buffer RLT, MN RA1 Macherey–Nagel NucleoSpin Plant using buffer RA1, ZR ZR plant RNA MiniPrep, OPR Omega EZNA plant RNA Kit 1, QRU Qiagen RNeasy plus universal

    Techniques Used: Modification

    28) Product Images from "Molecular Cloning and Characterization of Novel Phytocystatin Gene from Turmeric, Curcuma longa"

    Article Title: Molecular Cloning and Characterization of Novel Phytocystatin Gene from Turmeric, Curcuma longa

    Journal: BioMed Research International

    doi: 10.1155/2014/973790

    Total RNA integrity test on 1.0% agarose gel. Intact 28S and 18S total RNA bands can be observed on the agarose gel, indicating a good integrity of the total RNA after being extracted by different methods (lanes 1 and 2: modified CTAB method, lane 3, 4: RNAzol RT, and lane 5, 6: RNeasy Plant Mini kit).
    Figure Legend Snippet: Total RNA integrity test on 1.0% agarose gel. Intact 28S and 18S total RNA bands can be observed on the agarose gel, indicating a good integrity of the total RNA after being extracted by different methods (lanes 1 and 2: modified CTAB method, lane 3, 4: RNAzol RT, and lane 5, 6: RNeasy Plant Mini kit).

    Techniques Used: Agarose Gel Electrophoresis, Modification

    29) Product Images from "Filter paper-based spin column method for cost-efficient DNA or RNA purification"

    Article Title: Filter paper-based spin column method for cost-efficient DNA or RNA purification

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0203011

    The efficiency of filter paper for purification of nucleic acids from various sources using respective Qiagen kits. (A) Tomato genomic DNAs purified using Qiagen DNeasy plant mini kit. (B) Tomato total RNAs purified using Qiagen RNeasy plant mini kit. (C) PCR products of a GUS fragment purified using Qiagen QIAquick PCR purification kit. (D) PCR products of GUS fragment recovered from an agarose gel using a Qiagen QIAquick gel extraction kit. (E) pUC -19 plasmid DNAs purified using a Qiagen QIAprep spin miniprep kit. For each panel, from left to right are (Q) nucleic acid purified in experiments using original Qiagen spin column, (G) reassembled spin column using two layers of Whatman glass microfiber filters (Grade GF/F), and (P) reassembled spin column using two layers of Whatman qualitative filter paper, (Grade 3) respectively. Upper panel is quantification data based on three experimental replicates normalized according to performance of the Qiagen kit; lower panel is an image of agarose gel electrophoresis for the same volume of purified nucleic acids.
    Figure Legend Snippet: The efficiency of filter paper for purification of nucleic acids from various sources using respective Qiagen kits. (A) Tomato genomic DNAs purified using Qiagen DNeasy plant mini kit. (B) Tomato total RNAs purified using Qiagen RNeasy plant mini kit. (C) PCR products of a GUS fragment purified using Qiagen QIAquick PCR purification kit. (D) PCR products of GUS fragment recovered from an agarose gel using a Qiagen QIAquick gel extraction kit. (E) pUC -19 plasmid DNAs purified using a Qiagen QIAprep spin miniprep kit. For each panel, from left to right are (Q) nucleic acid purified in experiments using original Qiagen spin column, (G) reassembled spin column using two layers of Whatman glass microfiber filters (Grade GF/F), and (P) reassembled spin column using two layers of Whatman qualitative filter paper, (Grade 3) respectively. Upper panel is quantification data based on three experimental replicates normalized according to performance of the Qiagen kit; lower panel is an image of agarose gel electrophoresis for the same volume of purified nucleic acids.

    Techniques Used: Purification, Polymerase Chain Reaction, Agarose Gel Electrophoresis, Gel Extraction, Plasmid Preparation

    30) Product Images from "Molecular Cloning and Characterization of Novel Phytocystatin Gene from Turmeric, Curcuma longa"

    Article Title: Molecular Cloning and Characterization of Novel Phytocystatin Gene from Turmeric, Curcuma longa

    Journal: BioMed Research International

    doi: 10.1155/2014/973790

    Total RNA integrity test on 1.0% agarose gel. Intact 28S and 18S total RNA bands can be observed on the agarose gel, indicating a good integrity of the total RNA after being extracted by different methods (lanes 1 and 2: modified CTAB method, lane 3, 4: RNAzol RT, and lane 5, 6: RNeasy Plant Mini kit).
    Figure Legend Snippet: Total RNA integrity test on 1.0% agarose gel. Intact 28S and 18S total RNA bands can be observed on the agarose gel, indicating a good integrity of the total RNA after being extracted by different methods (lanes 1 and 2: modified CTAB method, lane 3, 4: RNAzol RT, and lane 5, 6: RNeasy Plant Mini kit).

    Techniques Used: Agarose Gel Electrophoresis, Modification

    31) Product Images from "Molecular Cloning and Characterization of Novel Phytocystatin Gene from Turmeric, Curcuma longa"

    Article Title: Molecular Cloning and Characterization of Novel Phytocystatin Gene from Turmeric, Curcuma longa

    Journal: BioMed Research International

    doi: 10.1155/2014/973790

    Total RNA integrity test on 1.0% agarose gel. Intact 28S and 18S total RNA bands can be observed on the agarose gel, indicating a good integrity of the total RNA after being extracted by different methods (lanes 1 and 2: modified CTAB method, lane 3, 4: RNAzol RT, and lane 5, 6: RNeasy Plant Mini kit).
    Figure Legend Snippet: Total RNA integrity test on 1.0% agarose gel. Intact 28S and 18S total RNA bands can be observed on the agarose gel, indicating a good integrity of the total RNA after being extracted by different methods (lanes 1 and 2: modified CTAB method, lane 3, 4: RNAzol RT, and lane 5, 6: RNeasy Plant Mini kit).

    Techniques Used: Agarose Gel Electrophoresis, Modification

    Related Articles

    Centrifugation:

    Article Title: Multiplex Detection of Aspergillus fumigatus Mycoviruses
    Article Snippet: .. RNA Extraction and Reverse Transcription In order to perform multiplex PCR for mycovirus-infected A. fumigatus isolates, total fungal RNA was extracted using the RNeasy Plant Mini kit (Qiagen, Hilden, Germany) from 100 mg of grounded mycelium, quantified by using NanoDrop 2000C spectrophotometer (Thermo Fischer, Waltham, MA, USA) and cDNA was synthesized using Superscript-III first-strand synthesis system (Invitrogen, Carlsbad, CA, USA) according to the manufacturers’ protocol as follows; 5 µg of RNA, 10 mM of dNTP mix and 250 ng of random primers along with 6.5 µL of DEPC-treated water were incubated at 65 °C for 5 min and snap cooled on ice for at least 2 min. Then 4 µL of 5× first strand buffer, 0.1 M DTT, 40 U of RNasin RNase inhibitor (Promega, Madison, WI, USA) and 200 U of Superscript-III reverse transcriptase were added to the reaction after a brief centrifugation. .. The 20 µL reaction mixture was then subjected to the following cycling regime of incubation at 25 °C, 50 °C and 70 °C for 5 min, 1 h and 15 min, respectively in a DNA Engine DYAD thermocycler.

    Amplification:

    Article Title: Multiplex Detection of Aspergillus fumigatus Mycoviruses
    Article Snippet: The Efficiency of the RNA Extraction Method The multiplex RT-PCR amplification assay was used to detect known and characterized A. fumigatus mycoviruses, namely AfuCV, AfuPV-1 and AfuTmV-1. .. The effect of the dsRNA extraction method on the efficiency of PCR was tested using LiCl extraction and the RNeasy Plant Mini kit (Qiagen).

    Article Title: Multiplex Detection of Aspergillus fumigatus Mycoviruses
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    Article Title: A Key ABA Catabolic Gene, OsABA8ox3, Is Involved in Drought Stress Resistance in Rice
    Article Snippet: Gene expression analysis by quantitative real-time PCR (qRT-PCR) Total RNA was extracted from rice seedlings using an RNA Easy Plant Mini Kit (Qiagen, CA) and then digested with DNase I (Amersham, USA) to eliminate genomic DNA contamination. .. The data was normalized to the amplification of rice ACTIN2 gene.

    Positive Control:

    Article Title: Multiplex Detection of Aspergillus fumigatus Mycoviruses
    Article Snippet: RNA Extraction and Reverse Transcription In order to perform multiplex PCR for mycovirus-infected A. fumigatus isolates, total fungal RNA was extracted using the RNeasy Plant Mini kit (Qiagen, Hilden, Germany) from 100 mg of grounded mycelium, quantified by using NanoDrop 2000C spectrophotometer (Thermo Fischer, Waltham, MA, USA) and cDNA was synthesized using Superscript-III first-strand synthesis system (Invitrogen, Carlsbad, CA, USA) according to the manufacturers’ protocol as follows; 5 µg of RNA, 10 mM of dNTP mix and 250 ng of random primers along with 6.5 µL of DEPC-treated water were incubated at 65 °C for 5 min and snap cooled on ice for at least 2 min. Then 4 µL of 5× first strand buffer, 0.1 M DTT, 40 U of RNasin RNase inhibitor (Promega, Madison, WI, USA) and 200 U of Superscript-III reverse transcriptase were added to the reaction after a brief centrifugation. .. Additionally, viral dsRNAs were extracted using LiCl extraction as described previously [ ] and used as a positive control in order to check the efficiency of the kit at extracting viral dsRNAs.

    Synthesized:

    Article Title: Multiplex Detection of Aspergillus fumigatus Mycoviruses
    Article Snippet: .. RNA Extraction and Reverse Transcription In order to perform multiplex PCR for mycovirus-infected A. fumigatus isolates, total fungal RNA was extracted using the RNeasy Plant Mini kit (Qiagen, Hilden, Germany) from 100 mg of grounded mycelium, quantified by using NanoDrop 2000C spectrophotometer (Thermo Fischer, Waltham, MA, USA) and cDNA was synthesized using Superscript-III first-strand synthesis system (Invitrogen, Carlsbad, CA, USA) according to the manufacturers’ protocol as follows; 5 µg of RNA, 10 mM of dNTP mix and 250 ng of random primers along with 6.5 µL of DEPC-treated water were incubated at 65 °C for 5 min and snap cooled on ice for at least 2 min. Then 4 µL of 5× first strand buffer, 0.1 M DTT, 40 U of RNasin RNase inhibitor (Promega, Madison, WI, USA) and 200 U of Superscript-III reverse transcriptase were added to the reaction after a brief centrifugation. .. The 20 µL reaction mixture was then subjected to the following cycling regime of incubation at 25 °C, 50 °C and 70 °C for 5 min, 1 h and 15 min, respectively in a DNA Engine DYAD thermocycler.

    Article Title: A Key ABA Catabolic Gene, OsABA8ox3, Is Involved in Drought Stress Resistance in Rice
    Article Snippet: Gene expression analysis by quantitative real-time PCR (qRT-PCR) Total RNA was extracted from rice seedlings using an RNA Easy Plant Mini Kit (Qiagen, CA) and then digested with DNase I (Amersham, USA) to eliminate genomic DNA contamination. .. First-strand cDNA was synthesized using a SuperScript first-strand synthesis system (Invitrogen, USA).

    Quantitative RT-PCR:

    Article Title: A Key ABA Catabolic Gene, OsABA8ox3, Is Involved in Drought Stress Resistance in Rice
    Article Snippet: .. Gene expression analysis by quantitative real-time PCR (qRT-PCR) Total RNA was extracted from rice seedlings using an RNA Easy Plant Mini Kit (Qiagen, CA) and then digested with DNase I (Amersham, USA) to eliminate genomic DNA contamination. .. First-strand cDNA was synthesized using a SuperScript first-strand synthesis system (Invitrogen, USA).

    SYBR Green Assay:

    Article Title: A Key ABA Catabolic Gene, OsABA8ox3, Is Involved in Drought Stress Resistance in Rice
    Article Snippet: Gene expression analysis by quantitative real-time PCR (qRT-PCR) Total RNA was extracted from rice seedlings using an RNA Easy Plant Mini Kit (Qiagen, CA) and then digested with DNase I (Amersham, USA) to eliminate genomic DNA contamination. .. Transcript levels of selected genes were measured by qRT-PCR using a iCycler (Bio-Rad, USA) with iQ SYBR Green Supermix (Bio-Rad, USA).

    Incubation:

    Article Title: Multiplex Detection of Aspergillus fumigatus Mycoviruses
    Article Snippet: .. RNA Extraction and Reverse Transcription In order to perform multiplex PCR for mycovirus-infected A. fumigatus isolates, total fungal RNA was extracted using the RNeasy Plant Mini kit (Qiagen, Hilden, Germany) from 100 mg of grounded mycelium, quantified by using NanoDrop 2000C spectrophotometer (Thermo Fischer, Waltham, MA, USA) and cDNA was synthesized using Superscript-III first-strand synthesis system (Invitrogen, Carlsbad, CA, USA) according to the manufacturers’ protocol as follows; 5 µg of RNA, 10 mM of dNTP mix and 250 ng of random primers along with 6.5 µL of DEPC-treated water were incubated at 65 °C for 5 min and snap cooled on ice for at least 2 min. Then 4 µL of 5× first strand buffer, 0.1 M DTT, 40 U of RNasin RNase inhibitor (Promega, Madison, WI, USA) and 200 U of Superscript-III reverse transcriptase were added to the reaction after a brief centrifugation. .. The 20 µL reaction mixture was then subjected to the following cycling regime of incubation at 25 °C, 50 °C and 70 °C for 5 min, 1 h and 15 min, respectively in a DNA Engine DYAD thermocycler.

    Article Title: Filter paper-based spin column method for cost-efficient DNA or RNA purification
    Article Snippet: Purification of plant RNA Plant total RNAs were purified using filter paper-based spin columns following the protocol of the Qiagen RNeasy Plant mini kit (RNeasy Mini Handbook. .. The tube with lysis mixture was incubated at 55°C for 1 to 3 min and mixed by inverting 2 to 3 times during incubation, and then centrifuged in a microcentrifuge at top speed (≥ 16,000 × g) for 10 mins to clear the lysate.

    Expressing:

    Article Title: Techniques for the Isolation of High-Quality RNA from Cells Encapsulated in Chitosan Hydrogels
    Article Snippet: In developing these regenerative approaches, gene expression analysis of the encapsulated cell populations by reverse transcriptase–polymerase chain reaction (RT-PCR) can provide useful information in characterizing the cellular response within the engineered microenvironments. .. Commercially available RNA extraction kits such as the Qiagen RNeasy® Mini Plant kit or the RNeasy® Mini kit have also been used as methods for extracting RNA from agarose and gellan, as well as alginate-based scaffolds., In general, most of these methods initially involve a form of mechanical disruption to help separate the RNA.

    Article Title: A Key ABA Catabolic Gene, OsABA8ox3, Is Involved in Drought Stress Resistance in Rice
    Article Snippet: .. Gene expression analysis by quantitative real-time PCR (qRT-PCR) Total RNA was extracted from rice seedlings using an RNA Easy Plant Mini Kit (Qiagen, CA) and then digested with DNase I (Amersham, USA) to eliminate genomic DNA contamination. .. First-strand cDNA was synthesized using a SuperScript first-strand synthesis system (Invitrogen, USA).

    Modification:

    Article Title: A method for extracting high-quality total RNA from plant rich in polysaccharides and polyphenols using Dendrobium huoshanense
    Article Snippet: In this study, D . huoshanense was selected as the subject to extract total RNA from its stem, leaf and flower by applying modified CHAN method [ – ]. .. In addition, Trizol (Lot no. 135404, Invitrogen, USA), RNeasy Plant Mini Kit (Lot no. 154048665, Qiagen, Germany) and RNAprep Pure Plant Kit (Lot no. Q5510, Tiangen, China) were also used to provide more data for the comparison regarding efficiency of different approaches.

    Article Title: Molecular Cloning and Characterization of Novel Phytocystatin Gene from Turmeric, Curcuma longa
    Article Snippet: .. Comparison of Total RNA Extraction with Different Methods Total RNA of turmeric (Curcuma longa) was successfully extracted from mature leaves of turmeric plant by three different methods: (i) modified CTAB (cetyltrimethyl ammonium bromide) method, (ii) RNAzol RT (Molecular Research Center Inc., USA), and (iii) RNeasy Plant Mini kit (Qiagen, USA). .. Intact bands of the 28S and 18S RNA can be clearly observed in the agarose gel for all the three methods ( ).

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Multiplex Detection of Aspergillus fumigatus Mycoviruses
    Article Snippet: The Efficiency of the RNA Extraction Method The multiplex RT-PCR amplification assay was used to detect known and characterized A. fumigatus mycoviruses, namely AfuCV, AfuPV-1 and AfuTmV-1. .. The effect of the dsRNA extraction method on the efficiency of PCR was tested using LiCl extraction and the RNeasy Plant Mini kit (Qiagen).

    Article Title: Techniques for the Isolation of High-Quality RNA from Cells Encapsulated in Chitosan Hydrogels
    Article Snippet: In developing these regenerative approaches, gene expression analysis of the encapsulated cell populations by reverse transcriptase–polymerase chain reaction (RT-PCR) can provide useful information in characterizing the cellular response within the engineered microenvironments. .. Commercially available RNA extraction kits such as the Qiagen RNeasy® Mini Plant kit or the RNeasy® Mini kit have also been used as methods for extracting RNA from agarose and gellan, as well as alginate-based scaffolds., In general, most of these methods initially involve a form of mechanical disruption to help separate the RNA.

    In Vivo:

    Article Title: Techniques for the Isolation of High-Quality RNA from Cells Encapsulated in Chitosan Hydrogels
    Article Snippet: Commercially available RNA extraction kits such as the Qiagen RNeasy® Mini Plant kit or the RNeasy® Mini kit have also been used as methods for extracting RNA from agarose and gellan, as well as alginate-based scaffolds., In general, most of these methods initially involve a form of mechanical disruption to help separate the RNA. .. Moreover, in vivo studies have shown that chitosan scaffolds are degraded primarily by lysozyme to produce oligosaccharide products.

    RNA Sequencing Assay:

    Article Title: Efficient and High-Quality RNA Isolation from Metabolite-Rich Tissues of Stevia rebaudiana, an Important Commercial Crop
    Article Snippet: Comparison with Previously-Reported RNA Extraction Methods from Stevia rebaudiana Commercially-available kits, such as Qiagen RNA plant mini kit (Qiagen, Hilden, Germany) and Spectrum Plant Total RNA Kit (Sigma-Aldrich, Missouri, USA), had been successfully used to isolate RNA from the leaf tissues of Stevia rebaudiana ( ; ). .. The RIN numbers of the RNA samples were fairly high (RIN > 7), allowing these samples to be used for library preparation and RNA-Seq.

    Isolation:

    Article Title: A highly effective and versatile technology for the isolation of RNAs from grapevines and other woody perennials for use in virus diagnostics
    Article Snippet: .. Commercial RNA isolation kits Five commercial kits were selected for this study, which included TRIzol Reagent (Life Technologies), RNeasy Plant mini kit (Qiagen), Spectrum™ Plant Total RNA kit (Sigma), AccuPrep viral RNA extraction kit (Bioneer) and Plant/fungi total RNA kit (Norgen BioTek). .. Methods of sample homogenization and RNA isolation

    Article Title: Subcellular Localization and Rolling Circle Replication of Peach Latent Mosaic Viroid: Hallmarks of Group A Viroids
    Article Snippet: Paragraph title: RNA isolation. ... The second procedure used the RNeasy Plant mini kit (Qiagen) to prepare RNA from ∼150 mg of tissue as specified by the manufacturer.

    Article Title: A highly effective and versatile technology for the isolation of RNAs from grapevines and other woody perennials for use in virus diagnostics
    Article Snippet: Comparison of five commercial kits in isolating RNA from peach and grapevine Five commonly used commercial RNA isolation kits were compared for their effectiveness in isolation of RNA from woody plants. .. These five kits are TRIzol Reagent (Life Technologies), RNeasy Plant mini kit (Qiagen), Spectrum™ Plant Total RNA kit (Sigma), AccuPrep viral RNA extraction kit (Bioneer) and Plant/fungi total RNA kit (Norgen BioTek).

    Article Title: Techniques for the Isolation of High-Quality RNA from Cells Encapsulated in Chitosan Hydrogels
    Article Snippet: However, the polysaccharide moieties within the scaffold materials can interfere with the RNA isolation process. .. Commercially available RNA extraction kits such as the Qiagen RNeasy® Mini Plant kit or the RNeasy® Mini kit have also been used as methods for extracting RNA from agarose and gellan, as well as alginate-based scaffolds., In general, most of these methods initially involve a form of mechanical disruption to help separate the RNA.

    Multiplex Assay:

    Article Title: Multiplex Detection of Aspergillus fumigatus Mycoviruses
    Article Snippet: The Efficiency of the RNA Extraction Method The multiplex RT-PCR amplification assay was used to detect known and characterized A. fumigatus mycoviruses, namely AfuCV, AfuPV-1 and AfuTmV-1. .. The effect of the dsRNA extraction method on the efficiency of PCR was tested using LiCl extraction and the RNeasy Plant Mini kit (Qiagen).

    Article Title: Multiplex Detection of Aspergillus fumigatus Mycoviruses
    Article Snippet: .. RNA Extraction and Reverse Transcription In order to perform multiplex PCR for mycovirus-infected A. fumigatus isolates, total fungal RNA was extracted using the RNeasy Plant Mini kit (Qiagen, Hilden, Germany) from 100 mg of grounded mycelium, quantified by using NanoDrop 2000C spectrophotometer (Thermo Fischer, Waltham, MA, USA) and cDNA was synthesized using Superscript-III first-strand synthesis system (Invitrogen, Carlsbad, CA, USA) according to the manufacturers’ protocol as follows; 5 µg of RNA, 10 mM of dNTP mix and 250 ng of random primers along with 6.5 µL of DEPC-treated water were incubated at 65 °C for 5 min and snap cooled on ice for at least 2 min. Then 4 µL of 5× first strand buffer, 0.1 M DTT, 40 U of RNasin RNase inhibitor (Promega, Madison, WI, USA) and 200 U of Superscript-III reverse transcriptase were added to the reaction after a brief centrifugation. .. The 20 µL reaction mixture was then subjected to the following cycling regime of incubation at 25 °C, 50 °C and 70 °C for 5 min, 1 h and 15 min, respectively in a DNA Engine DYAD thermocycler.

    Purification:

    Article Title: A method for extracting high-quality total RNA from plant rich in polysaccharides and polyphenols using Dendrobium huoshanense
    Article Snippet: However, due to similarity in physical and chemical properties between RNA and polysaccharides, it can coprecipitate with RNA during nucleotide purification process [ ]. .. In addition, Trizol (Lot no. 135404, Invitrogen, USA), RNeasy Plant Mini Kit (Lot no. 154048665, Qiagen, Germany) and RNAprep Pure Plant Kit (Lot no. Q5510, Tiangen, China) were also used to provide more data for the comparison regarding efficiency of different approaches.

    Article Title: Multiplex Detection of Aspergillus fumigatus Mycoviruses
    Article Snippet: The effect of the dsRNA extraction method on the efficiency of PCR was tested using LiCl extraction and the RNeasy Plant Mini kit (Qiagen). .. We demonstrated that targets of interest can be amplified with the same efficiency from viral dsRNAs obtained using different procedures such as virus purification or an RNA extraction kit.

    Article Title: Filter paper-based spin column method for cost-efficient DNA or RNA purification
    Article Snippet: .. Purification of plant RNA Plant total RNAs were purified using filter paper-based spin columns following the protocol of the Qiagen RNeasy Plant mini kit (RNeasy Mini Handbook. ..

    Polymerase Chain Reaction:

    Article Title: Multiplex Detection of Aspergillus fumigatus Mycoviruses
    Article Snippet: .. The effect of the dsRNA extraction method on the efficiency of PCR was tested using LiCl extraction and the RNeasy Plant Mini kit (Qiagen). ..

    Article Title: Multiplex Detection of Aspergillus fumigatus Mycoviruses
    Article Snippet: .. RNA Extraction and Reverse Transcription In order to perform multiplex PCR for mycovirus-infected A. fumigatus isolates, total fungal RNA was extracted using the RNeasy Plant Mini kit (Qiagen, Hilden, Germany) from 100 mg of grounded mycelium, quantified by using NanoDrop 2000C spectrophotometer (Thermo Fischer, Waltham, MA, USA) and cDNA was synthesized using Superscript-III first-strand synthesis system (Invitrogen, Carlsbad, CA, USA) according to the manufacturers’ protocol as follows; 5 µg of RNA, 10 mM of dNTP mix and 250 ng of random primers along with 6.5 µL of DEPC-treated water were incubated at 65 °C for 5 min and snap cooled on ice for at least 2 min. Then 4 µL of 5× first strand buffer, 0.1 M DTT, 40 U of RNasin RNase inhibitor (Promega, Madison, WI, USA) and 200 U of Superscript-III reverse transcriptase were added to the reaction after a brief centrifugation. .. The 20 µL reaction mixture was then subjected to the following cycling regime of incubation at 25 °C, 50 °C and 70 °C for 5 min, 1 h and 15 min, respectively in a DNA Engine DYAD thermocycler.

    Real-time Polymerase Chain Reaction:

    Article Title: A Key ABA Catabolic Gene, OsABA8ox3, Is Involved in Drought Stress Resistance in Rice
    Article Snippet: .. Gene expression analysis by quantitative real-time PCR (qRT-PCR) Total RNA was extracted from rice seedlings using an RNA Easy Plant Mini Kit (Qiagen, CA) and then digested with DNase I (Amersham, USA) to eliminate genomic DNA contamination. .. First-strand cDNA was synthesized using a SuperScript first-strand synthesis system (Invitrogen, USA).

    RNA Extraction:

    Article Title: A highly effective and versatile technology for the isolation of RNAs from grapevines and other woody perennials for use in virus diagnostics
    Article Snippet: .. Commercial RNA isolation kits Five commercial kits were selected for this study, which included TRIzol Reagent (Life Technologies), RNeasy Plant mini kit (Qiagen), Spectrum™ Plant Total RNA kit (Sigma), AccuPrep viral RNA extraction kit (Bioneer) and Plant/fungi total RNA kit (Norgen BioTek). .. Methods of sample homogenization and RNA isolation

    Article Title: A method for extracting high-quality total RNA from plant rich in polysaccharides and polyphenols using Dendrobium huoshanense
    Article Snippet: In addition, Trizol (Lot no. 135404, Invitrogen, USA), RNeasy Plant Mini Kit (Lot no. 154048665, Qiagen, Germany) and RNAprep Pure Plant Kit (Lot no. Q5510, Tiangen, China) were also used to provide more data for the comparison regarding efficiency of different approaches. .. Here, we provide a simple, economical and efficient RNA extraction method for plants rich in polysaccharides and polyphenols.

    Article Title: Efficient and High-Quality RNA Isolation from Metabolite-Rich Tissues of Stevia rebaudiana, an Important Commercial Crop
    Article Snippet: .. Comparison with Previously-Reported RNA Extraction Methods from Stevia rebaudiana Commercially-available kits, such as Qiagen RNA plant mini kit (Qiagen, Hilden, Germany) and Spectrum Plant Total RNA Kit (Sigma-Aldrich, Missouri, USA), had been successfully used to isolate RNA from the leaf tissues of Stevia rebaudiana ( ; ). .. The RIN numbers of the RNA samples were fairly high (RIN > 7), allowing these samples to be used for library preparation and RNA-Seq.

    Article Title: Multiplex Detection of Aspergillus fumigatus Mycoviruses
    Article Snippet: Paragraph title: 3.1. The Efficiency of the RNA Extraction Method ... The effect of the dsRNA extraction method on the efficiency of PCR was tested using LiCl extraction and the RNeasy Plant Mini kit (Qiagen).

    Article Title: Multiplex Detection of Aspergillus fumigatus Mycoviruses
    Article Snippet: .. RNA Extraction and Reverse Transcription In order to perform multiplex PCR for mycovirus-infected A. fumigatus isolates, total fungal RNA was extracted using the RNeasy Plant Mini kit (Qiagen, Hilden, Germany) from 100 mg of grounded mycelium, quantified by using NanoDrop 2000C spectrophotometer (Thermo Fischer, Waltham, MA, USA) and cDNA was synthesized using Superscript-III first-strand synthesis system (Invitrogen, Carlsbad, CA, USA) according to the manufacturers’ protocol as follows; 5 µg of RNA, 10 mM of dNTP mix and 250 ng of random primers along with 6.5 µL of DEPC-treated water were incubated at 65 °C for 5 min and snap cooled on ice for at least 2 min. Then 4 µL of 5× first strand buffer, 0.1 M DTT, 40 U of RNasin RNase inhibitor (Promega, Madison, WI, USA) and 200 U of Superscript-III reverse transcriptase were added to the reaction after a brief centrifugation. .. The 20 µL reaction mixture was then subjected to the following cycling regime of incubation at 25 °C, 50 °C and 70 °C for 5 min, 1 h and 15 min, respectively in a DNA Engine DYAD thermocycler.

    Article Title: A highly effective and versatile technology for the isolation of RNAs from grapevines and other woody perennials for use in virus diagnostics
    Article Snippet: .. These five kits are TRIzol Reagent (Life Technologies), RNeasy Plant mini kit (Qiagen), Spectrum™ Plant Total RNA kit (Sigma), AccuPrep viral RNA extraction kit (Bioneer) and Plant/fungi total RNA kit (Norgen BioTek). .. These kits were chosen due to their advantages in certain aspects in RNA isolation.

    Article Title: Molecular Cloning and Characterization of Novel Phytocystatin Gene from Turmeric, Curcuma longa
    Article Snippet: .. Comparison of Total RNA Extraction with Different Methods Total RNA of turmeric (Curcuma longa) was successfully extracted from mature leaves of turmeric plant by three different methods: (i) modified CTAB (cetyltrimethyl ammonium bromide) method, (ii) RNAzol RT (Molecular Research Center Inc., USA), and (iii) RNeasy Plant Mini kit (Qiagen, USA). .. Intact bands of the 28S and 18S RNA can be clearly observed in the agarose gel for all the three methods ( ).

    Article Title: Techniques for the Isolation of High-Quality RNA from Cells Encapsulated in Chitosan Hydrogels
    Article Snippet: .. Commercially available RNA extraction kits such as the Qiagen RNeasy® Mini Plant kit or the RNeasy® Mini kit have also been used as methods for extracting RNA from agarose and gellan, as well as alginate-based scaffolds., In general, most of these methods initially involve a form of mechanical disruption to help separate the RNA. .. However, the use of enzymatic digestion to partially degrade the polysaccharide scaffolds has not yet been explored as a means of improving the efficiency of RNA extraction from the hydrogels.

    Agarose Gel Electrophoresis:

    Article Title: Molecular Cloning and Characterization of Novel Phytocystatin Gene from Turmeric, Curcuma longa
    Article Snippet: Comparison of Total RNA Extraction with Different Methods Total RNA of turmeric (Curcuma longa) was successfully extracted from mature leaves of turmeric plant by three different methods: (i) modified CTAB (cetyltrimethyl ammonium bromide) method, (ii) RNAzol RT (Molecular Research Center Inc., USA), and (iii) RNeasy Plant Mini kit (Qiagen, USA). .. Intact bands of the 28S and 18S RNA can be clearly observed in the agarose gel for all the three methods ( ).

    Spectrophotometry:

    Article Title: Multiplex Detection of Aspergillus fumigatus Mycoviruses
    Article Snippet: .. RNA Extraction and Reverse Transcription In order to perform multiplex PCR for mycovirus-infected A. fumigatus isolates, total fungal RNA was extracted using the RNeasy Plant Mini kit (Qiagen, Hilden, Germany) from 100 mg of grounded mycelium, quantified by using NanoDrop 2000C spectrophotometer (Thermo Fischer, Waltham, MA, USA) and cDNA was synthesized using Superscript-III first-strand synthesis system (Invitrogen, Carlsbad, CA, USA) according to the manufacturers’ protocol as follows; 5 µg of RNA, 10 mM of dNTP mix and 250 ng of random primers along with 6.5 µL of DEPC-treated water were incubated at 65 °C for 5 min and snap cooled on ice for at least 2 min. Then 4 µL of 5× first strand buffer, 0.1 M DTT, 40 U of RNasin RNase inhibitor (Promega, Madison, WI, USA) and 200 U of Superscript-III reverse transcriptase were added to the reaction after a brief centrifugation. .. The 20 µL reaction mixture was then subjected to the following cycling regime of incubation at 25 °C, 50 °C and 70 °C for 5 min, 1 h and 15 min, respectively in a DNA Engine DYAD thermocycler.

    Article Title: Subcellular Localization and Rolling Circle Replication of Peach Latent Mosaic Viroid: Hallmarks of Group A Viroids
    Article Snippet: The second procedure used the RNeasy Plant mini kit (Qiagen) to prepare RNA from ∼150 mg of tissue as specified by the manufacturer. .. All RNA samples were quantified by UV spectrophotometry and electrophoresed on 1.3% agarose gels to assess the quality of the preparation.

    Produced:

    Article Title: Molecular Cloning and Characterization of Novel Phytocystatin Gene from Turmeric, Curcuma longa
    Article Snippet: Comparison of Total RNA Extraction with Different Methods Total RNA of turmeric (Curcuma longa) was successfully extracted from mature leaves of turmeric plant by three different methods: (i) modified CTAB (cetyltrimethyl ammonium bromide) method, (ii) RNAzol RT (Molecular Research Center Inc., USA), and (iii) RNeasy Plant Mini kit (Qiagen, USA). .. As for the quantitative test of the total RNA, with the raw tissue weight of 200 mg, our results showed that total RNA extraction by RNAzol RT produced the highest yield with concentration of ~800 ng/μ L while extraction by RNeasy Plant Mini kit and by a modified CTAB method yields ~300 ng/μ L of total RNA.

    Concentration Assay:

    Article Title: Subcellular Localization and Rolling Circle Replication of Peach Latent Mosaic Viroid: Hallmarks of Group A Viroids
    Article Snippet: The resulting supernatant was transferred to a fresh tube, and powdered PEG 6000 was added to a final concentration of 20% (wt/vol). .. The second procedure used the RNeasy Plant mini kit (Qiagen) to prepare RNA from ∼150 mg of tissue as specified by the manufacturer.

    Article Title: Molecular Cloning and Characterization of Novel Phytocystatin Gene from Turmeric, Curcuma longa
    Article Snippet: Comparison of Total RNA Extraction with Different Methods Total RNA of turmeric (Curcuma longa) was successfully extracted from mature leaves of turmeric plant by three different methods: (i) modified CTAB (cetyltrimethyl ammonium bromide) method, (ii) RNAzol RT (Molecular Research Center Inc., USA), and (iii) RNeasy Plant Mini kit (Qiagen, USA). .. As for the quantitative test of the total RNA, with the raw tissue weight of 200 mg, our results showed that total RNA extraction by RNAzol RT produced the highest yield with concentration of ~800 ng/μ L while extraction by RNeasy Plant Mini kit and by a modified CTAB method yields ~300 ng/μ L of total RNA.

    Lysis:

    Article Title: Techniques for the Isolation of High-Quality RNA from Cells Encapsulated in Chitosan Hydrogels
    Article Snippet: Furthermore, RNA quality can be compromised during the cell lysis steps of the extraction in cationic scaffolds, such as those prepared from chitosan, as insoluble ionic complexes can form with existing soluble anions, including polysaccharides, glycosaminoglycans, and DNA fragments., Thus, conventional RNA extraction techniques used for two-dimensional cell cultures or whole tissues that rely on monophasic phenol and guanidine isothiocyanate solutions or the use of β-mercaptoethanol, N-laurosylsarcosine (sarkosyl), and density gradient centrifugation in cesium trifluoroacetate alone may not be adequate. .. Commercially available RNA extraction kits such as the Qiagen RNeasy® Mini Plant kit or the RNeasy® Mini kit have also been used as methods for extracting RNA from agarose and gellan, as well as alginate-based scaffolds., In general, most of these methods initially involve a form of mechanical disruption to help separate the RNA.

    Article Title: Filter paper-based spin column method for cost-efficient DNA or RNA purification
    Article Snippet: Purification of plant RNA Plant total RNAs were purified using filter paper-based spin columns following the protocol of the Qiagen RNeasy Plant mini kit (RNeasy Mini Handbook. .. Up to 100 mg of ground frozen sample collected in 1.5 or 2 ml tube was suspended in 450 μl RTL or RLC buffer (Qiagen kit) with 1% (v/v) β -mercaptoethanol or homemade lysis buffer (8 M guanidine hydrochloride, 20 mM MES and 20 mM EDTA) with 1% (v/v) β -mercaptoethanol.

    Gradient Centrifugation:

    Article Title: Techniques for the Isolation of High-Quality RNA from Cells Encapsulated in Chitosan Hydrogels
    Article Snippet: Furthermore, RNA quality can be compromised during the cell lysis steps of the extraction in cationic scaffolds, such as those prepared from chitosan, as insoluble ionic complexes can form with existing soluble anions, including polysaccharides, glycosaminoglycans, and DNA fragments., Thus, conventional RNA extraction techniques used for two-dimensional cell cultures or whole tissues that rely on monophasic phenol and guanidine isothiocyanate solutions or the use of β-mercaptoethanol, N-laurosylsarcosine (sarkosyl), and density gradient centrifugation in cesium trifluoroacetate alone may not be adequate. .. Commercially available RNA extraction kits such as the Qiagen RNeasy® Mini Plant kit or the RNeasy® Mini kit have also been used as methods for extracting RNA from agarose and gellan, as well as alginate-based scaffolds., In general, most of these methods initially involve a form of mechanical disruption to help separate the RNA.

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    Qiagen rneasy plant mini kit
    Conventional <t>PCR</t> to check amplicon size prior to multiplex PCR. Amplicon sizes were checked on two percent agarose gel prior to performing multiplex PCR for A. fumigatus dsRNA mycoviruses. AfuCV, AfuPV-1 and AfuTmV-1 dsRNAs were extracted using LiCl extraction (Lanes 2, 4 and 6, respectively) and <t>RNeasy</t> Plant Mini kit (Lanes 3, 5 and 7, respectively) and used as templates for amplification with AfuCV, AfuPV-1 and AfuTmV-1 primers (2–3, 4–5, 6–7, respectively). Hyperladder-I was used as a marker to estimate the size of the amplicons (Lane 1).
    Rneasy Plant Mini Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 1756 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Conventional PCR to check amplicon size prior to multiplex PCR. Amplicon sizes were checked on two percent agarose gel prior to performing multiplex PCR for A. fumigatus dsRNA mycoviruses. AfuCV, AfuPV-1 and AfuTmV-1 dsRNAs were extracted using LiCl extraction (Lanes 2, 4 and 6, respectively) and RNeasy Plant Mini kit (Lanes 3, 5 and 7, respectively) and used as templates for amplification with AfuCV, AfuPV-1 and AfuTmV-1 primers (2–3, 4–5, 6–7, respectively). Hyperladder-I was used as a marker to estimate the size of the amplicons (Lane 1).

    Journal: Viruses

    Article Title: Multiplex Detection of Aspergillus fumigatus Mycoviruses

    doi: 10.3390/v10050247

    Figure Lengend Snippet: Conventional PCR to check amplicon size prior to multiplex PCR. Amplicon sizes were checked on two percent agarose gel prior to performing multiplex PCR for A. fumigatus dsRNA mycoviruses. AfuCV, AfuPV-1 and AfuTmV-1 dsRNAs were extracted using LiCl extraction (Lanes 2, 4 and 6, respectively) and RNeasy Plant Mini kit (Lanes 3, 5 and 7, respectively) and used as templates for amplification with AfuCV, AfuPV-1 and AfuTmV-1 primers (2–3, 4–5, 6–7, respectively). Hyperladder-I was used as a marker to estimate the size of the amplicons (Lane 1).

    Article Snippet: The effect of the dsRNA extraction method on the efficiency of PCR was tested using LiCl extraction and the RNeasy Plant Mini kit (Qiagen).

    Techniques: Polymerase Chain Reaction, Amplification, Multiplex Assay, Agarose Gel Electrophoresis, Marker

    The efficiency of filter paper for purification of nucleic acids from various sources using respective Qiagen kits. (A) Tomato genomic DNAs purified using Qiagen DNeasy plant mini kit. (B) Tomato total RNAs purified using Qiagen RNeasy plant mini kit. (C) PCR products of a GUS fragment purified using Qiagen QIAquick PCR purification kit. (D) PCR products of GUS fragment recovered from an agarose gel using a Qiagen QIAquick gel extraction kit. (E) pUC -19 plasmid DNAs purified using a Qiagen QIAprep spin miniprep kit. For each panel, from left to right are (Q) nucleic acid purified in experiments using original Qiagen spin column, (G) reassembled spin column using two layers of Whatman glass microfiber filters (Grade GF/F), and (P) reassembled spin column using two layers of Whatman qualitative filter paper, (Grade 3) respectively. Upper panel is quantification data based on three experimental replicates normalized according to performance of the Qiagen kit; lower panel is an image of agarose gel electrophoresis for the same volume of purified nucleic acids.

    Journal: PLoS ONE

    Article Title: Filter paper-based spin column method for cost-efficient DNA or RNA purification

    doi: 10.1371/journal.pone.0203011

    Figure Lengend Snippet: The efficiency of filter paper for purification of nucleic acids from various sources using respective Qiagen kits. (A) Tomato genomic DNAs purified using Qiagen DNeasy plant mini kit. (B) Tomato total RNAs purified using Qiagen RNeasy plant mini kit. (C) PCR products of a GUS fragment purified using Qiagen QIAquick PCR purification kit. (D) PCR products of GUS fragment recovered from an agarose gel using a Qiagen QIAquick gel extraction kit. (E) pUC -19 plasmid DNAs purified using a Qiagen QIAprep spin miniprep kit. For each panel, from left to right are (Q) nucleic acid purified in experiments using original Qiagen spin column, (G) reassembled spin column using two layers of Whatman glass microfiber filters (Grade GF/F), and (P) reassembled spin column using two layers of Whatman qualitative filter paper, (Grade 3) respectively. Upper panel is quantification data based on three experimental replicates normalized according to performance of the Qiagen kit; lower panel is an image of agarose gel electrophoresis for the same volume of purified nucleic acids.

    Article Snippet: Purification of plant RNA Plant total RNAs were purified using filter paper-based spin columns following the protocol of the Qiagen RNeasy Plant mini kit (RNeasy Mini Handbook.

    Techniques: Purification, Polymerase Chain Reaction, Agarose Gel Electrophoresis, Gel Extraction, Plasmid Preparation

    Evaluation of purification of tobacco genomic DNA and total RNA using filter paper-based spin columns with respective Qiagen kit buffers and homemade buffers. (A) Agarose gel electrophoresis for 2.5 μl tobacco genomic DNAs elution from purification experiments using Qiagen DNeasy plant mini kit buffers with Qiagen original spin column (Lane Q/Q), filter paper recharged used spin column (Lane Q/R) and filter paper-based homemade spin column (Lane Q/H*), followed by tobacco genomic DNAs purified using homemade buffer with Qiagen original spin column (Lane H/Q), filter paper recharged used spin column (Lane H/R) and filter paper-based homemade spin column (Lane H/H*). (B) UV spectrum curve of tobacco DNAs purified using Qiagen kit (Q/Q, black curve), filter paper recharged spin columns with Qiagen kit buffers (Q/R, blue curve) or homemade buffers (H/R, red curve) from the same amount leaf tissue. Y-axis is UV absorbance, and X-axis is wavelength (nM). (C) Amplification plots for three duplicated qPCR reactions contain 20 ng DNA purified using Qiagen kit (Q/Q, Blue curves) or DNA purified from filter paper recharged spin column with homemade buffer (H/R, Red curves) respectively. The x-axis is PCR cycle numbers, Y-axis is the level of SYBR fluorescence, and the green line is an arbitrary threshold to determine the Cq value (the fractional cycle number at which amplification curve meet threshold level). (D) MOPS-formaldehyde denaturing agarose gel electrophoresis separated 5 μl RNA purified using Qiagen RNeasy plant mini kit buffers with a Qiagen original spin column (Lane Q/Q), filter paper recharged used spin column (Lane Q/R) and homemade filter paper-based spin column (Lane Q/H*), followed total tobacco RNAs purified by using homemade buffer with Qiagen original spin column (Lane H/Q), filter paper recharged used spin column (Lane H/R) and filter paper-based homemade spin column (Lane H/H*). (E) UV spectrum of tobacco total RNA purified using Qiagen kit (Q/Q, black curve), filter paper recharged spin column with Qiagen RNeasy plant mini kit buffers (Q/R, blue curve) or homemade buffers (H/R, red curve). Y-axis is UV absorbance, and the X-axis is wavelength. (F) Amplification plots of three duplicated qRT-PCR reactions for 2.5 ng RNA purified using Qiagen kit (Q/Q, Blue curves) or RNA purified using filter paper recharged spin column with homemade buffer (H/R, Red curves) respectively. Note: * The starting material amount is 100 mg tobacco leaf tissue for experiments using a Qiagen spin column or filter paper recharged spin column, and half amount of plant sample (50 mg) used for homemade spin column purification. All DNAs or RNAs were eluted using 100 ul elution solution.

    Journal: PLoS ONE

    Article Title: Filter paper-based spin column method for cost-efficient DNA or RNA purification

    doi: 10.1371/journal.pone.0203011

    Figure Lengend Snippet: Evaluation of purification of tobacco genomic DNA and total RNA using filter paper-based spin columns with respective Qiagen kit buffers and homemade buffers. (A) Agarose gel electrophoresis for 2.5 μl tobacco genomic DNAs elution from purification experiments using Qiagen DNeasy plant mini kit buffers with Qiagen original spin column (Lane Q/Q), filter paper recharged used spin column (Lane Q/R) and filter paper-based homemade spin column (Lane Q/H*), followed by tobacco genomic DNAs purified using homemade buffer with Qiagen original spin column (Lane H/Q), filter paper recharged used spin column (Lane H/R) and filter paper-based homemade spin column (Lane H/H*). (B) UV spectrum curve of tobacco DNAs purified using Qiagen kit (Q/Q, black curve), filter paper recharged spin columns with Qiagen kit buffers (Q/R, blue curve) or homemade buffers (H/R, red curve) from the same amount leaf tissue. Y-axis is UV absorbance, and X-axis is wavelength (nM). (C) Amplification plots for three duplicated qPCR reactions contain 20 ng DNA purified using Qiagen kit (Q/Q, Blue curves) or DNA purified from filter paper recharged spin column with homemade buffer (H/R, Red curves) respectively. The x-axis is PCR cycle numbers, Y-axis is the level of SYBR fluorescence, and the green line is an arbitrary threshold to determine the Cq value (the fractional cycle number at which amplification curve meet threshold level). (D) MOPS-formaldehyde denaturing agarose gel electrophoresis separated 5 μl RNA purified using Qiagen RNeasy plant mini kit buffers with a Qiagen original spin column (Lane Q/Q), filter paper recharged used spin column (Lane Q/R) and homemade filter paper-based spin column (Lane Q/H*), followed total tobacco RNAs purified by using homemade buffer with Qiagen original spin column (Lane H/Q), filter paper recharged used spin column (Lane H/R) and filter paper-based homemade spin column (Lane H/H*). (E) UV spectrum of tobacco total RNA purified using Qiagen kit (Q/Q, black curve), filter paper recharged spin column with Qiagen RNeasy plant mini kit buffers (Q/R, blue curve) or homemade buffers (H/R, red curve). Y-axis is UV absorbance, and the X-axis is wavelength. (F) Amplification plots of three duplicated qRT-PCR reactions for 2.5 ng RNA purified using Qiagen kit (Q/Q, Blue curves) or RNA purified using filter paper recharged spin column with homemade buffer (H/R, Red curves) respectively. Note: * The starting material amount is 100 mg tobacco leaf tissue for experiments using a Qiagen spin column or filter paper recharged spin column, and half amount of plant sample (50 mg) used for homemade spin column purification. All DNAs or RNAs were eluted using 100 ul elution solution.

    Article Snippet: Purification of plant RNA Plant total RNAs were purified using filter paper-based spin columns following the protocol of the Qiagen RNeasy Plant mini kit (RNeasy Mini Handbook.

    Techniques: Purification, Agarose Gel Electrophoresis, Amplification, Real-time Polymerase Chain Reaction, Polymerase Chain Reaction, Fluorescence, Quantitative RT-PCR

    Profile of total RNA isolated by using five commercial kits. a Denaturing gel electrophoresis of total RNA isolated from peach. b Denaturing gel electrophoresis of total RNA from grapevine leaves. 50 mg of young peach and grapevine leaves (indicated as Y), and mature (M) grapevine leaves was used in RNA isolation with Spectrum™ Plant Total RNA kit (Sigma), RNeasy Plant mini kit (Qiagen), Plant/fungi total RNA kit (Norgen), AccuPrep viral RNA extraction kit (Bioneer) and TRIzol Reagent (Life Technologies). The total RNA yield (μg), A260/A280 and A260/A230 ratios averaged from two replicates are given below each gel panel. 28S rRNA, 18S rRNA and small RNAs are indicated with arrows. c Capillary electrophoresis of total RNA with Agilent Bioanalyzer. One ml of each of the total RNA preparations isolated using these five systems was used for the analysis with an Agilent Bioanalyzer 2100 equipped with an RNA Nano chip

    Journal: Virology Journal

    Article Title: A highly effective and versatile technology for the isolation of RNAs from grapevines and other woody perennials for use in virus diagnostics

    doi: 10.1186/s12985-015-0376-3

    Figure Lengend Snippet: Profile of total RNA isolated by using five commercial kits. a Denaturing gel electrophoresis of total RNA isolated from peach. b Denaturing gel electrophoresis of total RNA from grapevine leaves. 50 mg of young peach and grapevine leaves (indicated as Y), and mature (M) grapevine leaves was used in RNA isolation with Spectrum™ Plant Total RNA kit (Sigma), RNeasy Plant mini kit (Qiagen), Plant/fungi total RNA kit (Norgen), AccuPrep viral RNA extraction kit (Bioneer) and TRIzol Reagent (Life Technologies). The total RNA yield (μg), A260/A280 and A260/A230 ratios averaged from two replicates are given below each gel panel. 28S rRNA, 18S rRNA and small RNAs are indicated with arrows. c Capillary electrophoresis of total RNA with Agilent Bioanalyzer. One ml of each of the total RNA preparations isolated using these five systems was used for the analysis with an Agilent Bioanalyzer 2100 equipped with an RNA Nano chip

    Article Snippet: These five kits are TRIzol Reagent (Life Technologies), RNeasy Plant mini kit (Qiagen), Spectrum™ Plant Total RNA kit (Sigma), AccuPrep viral RNA extraction kit (Bioneer) and Plant/fungi total RNA kit (Norgen BioTek).

    Techniques: Isolation, Nucleic Acid Electrophoresis, RNA Extraction, Electrophoresis, Chromatin Immunoprecipitation

    Total RNA integrity test on 1.0% agarose gel. Intact 28S and 18S total RNA bands can be observed on the agarose gel, indicating a good integrity of the total RNA after being extracted by different methods (lanes 1 and 2: modified CTAB method, lane 3, 4: RNAzol RT, and lane 5, 6: RNeasy Plant Mini kit).

    Journal: BioMed Research International

    Article Title: Molecular Cloning and Characterization of Novel Phytocystatin Gene from Turmeric, Curcuma longa

    doi: 10.1155/2014/973790

    Figure Lengend Snippet: Total RNA integrity test on 1.0% agarose gel. Intact 28S and 18S total RNA bands can be observed on the agarose gel, indicating a good integrity of the total RNA after being extracted by different methods (lanes 1 and 2: modified CTAB method, lane 3, 4: RNAzol RT, and lane 5, 6: RNeasy Plant Mini kit).

    Article Snippet: Comparison of Total RNA Extraction with Different Methods Total RNA of turmeric (Curcuma longa) was successfully extracted from mature leaves of turmeric plant by three different methods: (i) modified CTAB (cetyltrimethyl ammonium bromide) method, (ii) RNAzol RT (Molecular Research Center Inc., USA), and (iii) RNeasy Plant Mini kit (Qiagen, USA).

    Techniques: Agarose Gel Electrophoresis, Modification