rpe  (Qiagen)


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    Name:
    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
    Buy from Supplier


    Structured Review

    Qiagen rpe
    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/rpe/product/Qiagen
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rpe - by Bioz Stars, 2021-03
    97/100 stars

    Images

    1) 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

    Development of RNA extraction protocol . A : RNA extracted from Arabidopsis leaves. Lane 1, RNA extraction using Logemann et al.'s protocol [ 4 ]. Lanes 2 and 3, RNA obtained by transfer through a plasmid DNA extraction column (QIAprep Spin Miniprep Kit). Lanes 4 and 5, RNA obtained from transfer through an RNA collection column (RNeasy). EtOH, 96% ethanol; M, DNA 1-kb ladder. B : RNA yield following extraction in 8 M guanidine hydrochloride buffer and 0.5 volume of 96% EtOH and transfer through a plasmid DNA extraction column (QIAprep Spin Miniprep Kit), followed by 2-3 washes in: (1) RW1 × 2 and RPE (RNeasy Plant Mini Kit), (2) 3 M Na-acetate and 70-75% EtOH; (3) two washes with 96% EtOH; (4) 3 M Na-acetate and PB (QIAprep); (5) two washes with PB. RNA yield was measured spectrophotometrically.
    Figure Legend Snippet: Development of RNA extraction protocol . A : RNA extracted from Arabidopsis leaves. Lane 1, RNA extraction using Logemann et al.'s protocol [ 4 ]. Lanes 2 and 3, RNA obtained by transfer through a plasmid DNA extraction column (QIAprep Spin Miniprep Kit). Lanes 4 and 5, RNA obtained from transfer through an RNA collection column (RNeasy). EtOH, 96% ethanol; M, DNA 1-kb ladder. B : RNA yield following extraction in 8 M guanidine hydrochloride buffer and 0.5 volume of 96% EtOH and transfer through a plasmid DNA extraction column (QIAprep Spin Miniprep Kit), followed by 2-3 washes in: (1) RW1 × 2 and RPE (RNeasy Plant Mini Kit), (2) 3 M Na-acetate and 70-75% EtOH; (3) two washes with 96% EtOH; (4) 3 M Na-acetate and PB (QIAprep); (5) two washes with PB. RNA yield was measured spectrophotometrically.

    Techniques Used: RNA Extraction, Plasmid Preparation, DNA Extraction

    Related Articles

    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: 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.

    RNA Extraction:

    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: 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). .. 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 ( ).

    Multiplex Assay:

    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.

    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.

    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.

    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.

    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.

    Modification:

    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 ( ).

    Purification:

    Article Title: Filter paper-based spin column method for cost-efficient DNA or RNA purification
    Article Snippet: DNA elution was collected into collection tube through centrifugation at 6000 × g for 1 min. .. 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. ..

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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 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rneasy plant mini kit/product/Qiagen
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rneasy plant mini kit - by Bioz Stars, 2021-03
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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

    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

    Journal: BMC Research Notes

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

    doi: 10.1186/s13104-017-2564-2

    Figure Lengend 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

    Article Snippet: Kits with alternate buffers, such as the Macherey–Nagel NucleoSpin Plant and Qiagen RNeasy Plant kits, tended to produce better results using the alternate buffers (Table ), which make them attractive options compared to kits with no alternates.

    Techniques: Modification

    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