rna plant mini kit  (Qiagen)


Bioz Verified Symbol Qiagen is a verified supplier
Bioz Manufacturer Symbol Qiagen manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 95
    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 rna plant mini kit
    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/rna plant mini kit/product/Qiagen
    Average 95 stars, based on 7 article reviews
    Price from $9.99 to $1999.99
    rna plant mini kit - by Bioz Stars, 2020-02
    95/100 stars

    Images

    1) 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

    Related Articles

    Clone Assay:

    Article Title: Experimental Transmission of Pospiviroid Populations to Weed Species Characteristic of Potato and Hop Fields ▿
    Article Snippet: For the reverse transcription-PCR (RT-PCR) and real-time PCR, total RNA was isolated from 100 mg of plant leaf tissue by using CONCERT (plant RNA purification reagent; Invitrogen) following RNA purification and DNA cleavage on columns (RNeasy Plant Total RNA kit; QIAGEN, Germany). .. RT-PCR amplification for viroid detection and cloning was performed by using Titan One-Tube RT-PCR (Roche) including a high-fidelity Pwo polymerase (Roche Molecular Biochemicals).

    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: A practical examination of RNA isolation methods for European pear (Pyrus communis)
    Article Snippet: .. However, if yield is not a critical issue (e.g. cDNA synthesis for PCR amplification) or amplification and cleanup of pooled samples is part of a design, then the convenient Macherey–Nagel NuleoSpin Plant or Qiagen RNeasy Plant kit are attractive choices. .. The CTAB method is more scalable, though given input requirements for modern applications such as transcriptome sequencing (i.e. Illumina TruSeq Stranded mRNA— http://www.illumina.com ), preparations yielding as little as 25 ng of total RNA may be sufficient.

    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. .. In an attempt to confirm if there was direct amplification from DNA or not, cDNA synthesis was performed without the reverse transcriptase enzyme.

    Article Title: Experimental Transmission of Pospiviroid Populations to Weed Species Characteristic of Potato and Hop Fields ▿
    Article Snippet: For the reverse transcription-PCR (RT-PCR) and real-time PCR, total RNA was isolated from 100 mg of plant leaf tissue by using CONCERT (plant RNA purification reagent; Invitrogen) following RNA purification and DNA cleavage on columns (RNeasy Plant Total RNA kit; QIAGEN, Germany). .. RT-PCR amplification for viroid detection and cloning was performed by using Titan One-Tube RT-PCR (Roche) including a high-fidelity Pwo polymerase (Roche Molecular Biochemicals).

    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.

    SYBR Green Assay:

    Article Title: A lysin motif effector subverts chitin‐triggered immunity to facilitate arbuscular mycorrhizal symbiosis
    Article Snippet: RNA isolation and quantitative PCR For all experiments, total RNAs were isolated using a Qiagen plant RNA mini kit or Omega EZNA Plant RNA kit (Omega Bio‐Tek, Norcross, GA, USA) following the manufacturer’s instructions, including on‐column DNAse treatment. .. Quantitative PCR (qPCR) was done using an iQ SYBR Green Supermix (Bio‐Rad) on a Bio‐Rad CFX real‐time platform.

    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.

    Activity Assay:

    Article Title: A practical examination of RNA isolation methods for European pear (Pyrus communis)
    Article Snippet: We eliminated intensive preparation of spatulas and mortars and pestles by baking at 200 °C prior to use because native plant RNase activity [ ] renders these steps redundant due to the processing of tissues with liquid N2 and the strong denaturing conditions of lysis buffers. .. However, if yield is not a critical issue (e.g. cDNA synthesis for PCR amplification) or amplification and cleanup of pooled samples is part of a design, then the convenient Macherey–Nagel NuleoSpin Plant or Qiagen RNeasy Plant kit are attractive choices.

    Article Title: A practical examination of RNA isolation methods for European pear (Pyrus communis)
    Article Snippet: Many plant tissues are recalcitrant to RNA extraction for a host of reasons including abundant secondary metabolites and polysaccharides, tough cell walls that may be lignified, high levels of native RNase activity, and widely variable amounts of RNA [ ]. .. However, high quality transcriptome data that followed successful RNA isolation using the Qiagen RNeasy Plant Kit™ [ ] has been reported for pear (Pyrus communis ‘Bartlett’).

    Modification:

    Article Title: A practical examination of RNA isolation methods for European pear (Pyrus communis)
    Article Snippet: We included apple fruit because the Gapper et al. [ ] protocol, on which ours was based, was used successfully on apples and we wished to confirm our modified protocol was useful for apple tissues as well. .. Based upon the results of the tests in stored ‘d’Anjou’ fruit, the Macherey–Nagel Nucleospin Plant kit was selected for additional testing based on the observation that the low yielding preparations were more or less of equivalent quality to the next best Qiagen RNeasy Plant kit, yet yields were more consistent and higher with the Macherey–Nagel product for stored ‘d’Anjou’ cortical tissues.

    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: A practical examination of RNA isolation methods for European pear (Pyrus communis)
    Article Snippet: Our modified CTAB protocol routinely produced excellent quality RNA with RINs ≥8.3, A260/280 ≥1.67 and the best yields across all tissue types ranging from 0.5 to 2 µg from ~100 mg fresh weight tissue (Table ; Figs. , ). .. 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.

    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 ( ).

    Polymerase Chain Reaction:

    Article Title: A practical examination of RNA isolation methods for European pear (Pyrus communis)
    Article Snippet: .. However, if yield is not a critical issue (e.g. cDNA synthesis for PCR amplification) or amplification and cleanup of pooled samples is part of a design, then the convenient Macherey–Nagel NuleoSpin Plant or Qiagen RNeasy Plant kit are attractive choices. .. The CTAB method is more scalable, though given input requirements for modern applications such as transcriptome sequencing (i.e. Illumina TruSeq Stranded mRNA— http://www.illumina.com ), preparations yielding as little as 25 ng of total RNA may be sufficient.

    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.

    Sequencing:

    Article Title: A practical examination of RNA isolation methods for European pear (Pyrus communis)
    Article Snippet: However, if yield is not a critical issue (e.g. cDNA synthesis for PCR amplification) or amplification and cleanup of pooled samples is part of a design, then the convenient Macherey–Nagel NuleoSpin Plant or Qiagen RNeasy Plant kit are attractive choices. .. The CTAB method is more scalable, though given input requirements for modern applications such as transcriptome sequencing (i.e. Illumina TruSeq Stranded mRNA— http://www.illumina.com ), preparations yielding as little as 25 ng of total RNA may be sufficient.

    Article Title: Experimental Transmission of Pospiviroid Populations to Weed Species Characteristic of Potato and Hop Fields ▿
    Article Snippet: For the reverse transcription-PCR (RT-PCR) and real-time PCR, total RNA was isolated from 100 mg of plant leaf tissue by using CONCERT (plant RNA purification reagent; Invitrogen) following RNA purification and DNA cleavage on columns (RNeasy Plant Total RNA kit; QIAGEN, Germany). .. The nonspecific adenine in each primer (indicated by a small letter “a”) was designed to facilitate the cleavage of cDNA fragments; restriction sites encoded in the primers are underlined, and the positions are numbered corresponding to the respective viroid sequence.

    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: A practical examination of RNA isolation methods for European pear (Pyrus communis)
    Article Snippet: .. However, high quality transcriptome data that followed successful RNA isolation using the Qiagen RNeasy Plant Kit™ [ ] has been reported for pear (Pyrus communis ‘Bartlett’). ..

    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: Experimental Transmission of Pospiviroid Populations to Weed Species Characteristic of Potato and Hop Fields ▿
    Article Snippet: .. For the reverse transcription-PCR (RT-PCR) and real-time PCR, total RNA was isolated from 100 mg of plant leaf tissue by using CONCERT (plant RNA purification reagent; Invitrogen) following RNA purification and DNA cleavage on columns (RNeasy Plant Total RNA kit; QIAGEN, Germany). .. RT-PCR amplification for viroid detection and cloning was performed by using Titan One-Tube RT-PCR (Roche) including a high-fidelity Pwo polymerase (Roche Molecular Biochemicals).

    Article Title: A lysin motif effector subverts chitin‐triggered immunity to facilitate arbuscular mycorrhizal symbiosis
    Article Snippet: .. RNA isolation and quantitative PCR For all experiments, total RNAs were isolated using a Qiagen plant RNA mini kit or Omega EZNA Plant RNA kit (Omega Bio‐Tek, Norcross, GA, USA) following the manufacturer’s instructions, including on‐column DNAse treatment. .. Either 1 µg or 500 ng total RNA was used for reverse transcription using an iScript cDNA synthesis kit (Bio‐Rad).

    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: Experimental Transmission of Pospiviroid Populations to Weed Species Characteristic of Potato and Hop Fields ▿
    Article Snippet: .. For the reverse transcription-PCR (RT-PCR) and real-time PCR, total RNA was isolated from 100 mg of plant leaf tissue by using CONCERT (plant RNA purification reagent; Invitrogen) following RNA purification and DNA cleavage on columns (RNeasy Plant Total RNA kit; QIAGEN, Germany). .. RT-PCR amplification for viroid detection and cloning was performed by using Titan One-Tube RT-PCR (Roche) including a high-fidelity Pwo polymerase (Roche Molecular Biochemicals).

    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. ..

    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: Experimental Transmission of Pospiviroid Populations to Weed Species Characteristic of Potato and Hop Fields ▿
    Article Snippet: .. For the reverse transcription-PCR (RT-PCR) and real-time PCR, total RNA was isolated from 100 mg of plant leaf tissue by using CONCERT (plant RNA purification reagent; Invitrogen) following RNA purification and DNA cleavage on columns (RNeasy Plant Total RNA kit; QIAGEN, Germany). .. RT-PCR amplification for viroid detection and cloning was performed by using Titan One-Tube RT-PCR (Roche) including a high-fidelity Pwo polymerase (Roche Molecular Biochemicals).

    RNA Extraction:

    Article Title: A practical examination of RNA isolation methods for European pear (Pyrus communis)
    Article Snippet: Generally older tissues are more recalcitrant to RNA extraction than younger ones for a variety of reasons including presence of co-extracted secondary metabolites [ ] and in the case of climacteric fruit, increases in substances that tend to co-precipitate with RNA [ – ]. .. However, if yield is not a critical issue (e.g. cDNA synthesis for PCR amplification) or amplification and cleanup of pooled samples is part of a design, then the convenient Macherey–Nagel NuleoSpin Plant or Qiagen RNeasy Plant kit are attractive choices.

    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 ( ).

    Real-time Polymerase Chain Reaction:

    Article Title: Experimental Transmission of Pospiviroid Populations to Weed Species Characteristic of Potato and Hop Fields ▿
    Article Snippet: .. For the reverse transcription-PCR (RT-PCR) and real-time PCR, total RNA was isolated from 100 mg of plant leaf tissue by using CONCERT (plant RNA purification reagent; Invitrogen) following RNA purification and DNA cleavage on columns (RNeasy Plant Total RNA kit; QIAGEN, Germany). .. RT-PCR amplification for viroid detection and cloning was performed by using Titan One-Tube RT-PCR (Roche) including a high-fidelity Pwo polymerase (Roche Molecular Biochemicals).

    Article Title: A lysin motif effector subverts chitin‐triggered immunity to facilitate arbuscular mycorrhizal symbiosis
    Article Snippet: .. RNA isolation and quantitative PCR For all experiments, total RNAs were isolated using a Qiagen plant RNA mini kit or Omega EZNA Plant RNA kit (Omega Bio‐Tek, Norcross, GA, USA) following the manufacturer’s instructions, including on‐column DNAse treatment. .. Either 1 µg or 500 ng total RNA was used for reverse transcription using an iScript cDNA synthesis kit (Bio‐Rad).

    Functional Assay:

    Article Title: A practical examination of RNA isolation methods for European pear (Pyrus communis)
    Article Snippet: However, high quality transcriptome data that followed successful RNA isolation using the Qiagen RNeasy Plant Kit™ [ ] has been reported for pear (Pyrus communis ‘Bartlett’). .. To facilitate efforts for functional genomics of tree fruit towards improving postharvest fruit quality [ ], we present here a practical examination of pear fruit RNA isolation methods including several commercially available kits as well as a rapid and robust CTAB-based method.

    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.

    Sampling:

    Article Title: A practical examination of RNA isolation methods for European pear (Pyrus communis)
    Article Snippet: Results and discussion The sampling strategy was to evaluate the relatively recalcitrant tissues first (stored d’Anjou pear fruit), followed by less recalcitrant tissues (freshly harvested mature pear and apple fruit). .. Based upon the results of the tests in stored ‘d’Anjou’ fruit, the Macherey–Nagel Nucleospin Plant kit was selected for additional testing based on the observation that the low yielding preparations were more or less of equivalent quality to the next best Qiagen RNeasy Plant kit, yet yields were more consistent and higher with the Macherey–Nagel product for stored ‘d’Anjou’ cortical tissues.

    Produced:

    Article Title: A practical examination of RNA isolation methods for European pear (Pyrus communis)
    Article Snippet: The commercially available kits produced widely variable results, though some were comparable to our benchmark CTAB method. .. 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.

    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: 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: A practical examination of RNA isolation methods for European pear (Pyrus communis)
    Article Snippet: We eliminated intensive preparation of spatulas and mortars and pestles by baking at 200 °C prior to use because native plant RNase activity [ ] renders these steps redundant due to the processing of tissues with liquid N2 and the strong denaturing conditions of lysis buffers. .. However, if yield is not a critical issue (e.g. cDNA synthesis for PCR amplification) or amplification and cleanup of pooled samples is part of a design, then the convenient Macherey–Nagel NuleoSpin Plant or Qiagen RNeasy Plant kit are attractive choices.

    Article Title: A practical examination of RNA isolation methods for European pear (Pyrus communis)
    Article Snippet: This issue is especially common when using lysis buffers that contain chaotropic agents [ , ] such as those found broadly in commercially available kits. .. However, high quality transcriptome data that followed successful RNA isolation using the Qiagen RNeasy Plant Kit™ [ ] has been reported for pear (Pyrus communis ‘Bartlett’).

    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.

    Hood:

    Article Title: A practical examination of RNA isolation methods for European pear (Pyrus communis)
    Article Snippet: Importantly, our CTAB based protocol was time equivalent to the kits that were tested, though more equipment intensive (e.g. requiring a fume hood for chloroform handling, a refrigerated centrifuge and a water bath). .. However, if yield is not a critical issue (e.g. cDNA synthesis for PCR amplification) or amplification and cleanup of pooled samples is part of a design, then the convenient Macherey–Nagel NuleoSpin Plant or Qiagen RNeasy Plant kit are attractive choices.

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 95
    Qiagen rneasy plant mini kit
    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), <t>RNeasy</t> Plant mini kit (Qiagen), Plant/fungi total RNA kit (Norgen), AccuPrep viral RNA extraction kit (Bioneer) and <t>TRIzol</t> 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
    Rneasy Plant Mini Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 95/100, based on 1760 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rneasy plant mini kit/product/Qiagen
    Average 95 stars, based on 1760 article reviews
    Price from $9.99 to $1999.99
    rneasy plant mini kit - by Bioz Stars, 2020-02
    95/100 stars
      Buy from Supplier

    Image Search Results


    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

    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: 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.

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

    RiSLM ) is highly induced in intraradical mycelium (IRM) in a wide range of host plants. (a) RiSLM is highly induced in five hosts Medicago truncatula (Medicago), Allium schoenoprasum (Chives), Nicotiana benthamiana (Nicotiana), Solanum lycopersicum (Tomato) and Lotus japonicus (Lotus) compared with germinating spores (GS). Transcripts per million (TPM) was used to represent expression levels. Error bars represent SE from either three replicates (for Medicago, Chives, and Nicotiana) or two replicates (for Lotus and Tomato), collected from independent RNA sequencing (RNAseq) experiments. (b) Quantitative PCR analysis, showing that RiSLM (relative to Rhizophagus irregularis elongation factor 1α , RiEF ) is induced during arbuscular mychorrizal colonization and arbuscule (ARB) development as represented by relative expression of RiEF and the ARB‐specific phosphate transporter ( MtPT4 ) normalized by Medicago elongation factor 1α ( MtEF ). Error bars represent SE from three biological replicates. (c) RiSLM expression based on RNAseq analyses of laser microdissected ARBs and IRM compared with extraradical mycelium (ERM) and GS. Error bars represent SE from three biological replicates. For all figures, different characters indicate significant differences (least significant difference P

    Journal: The New Phytologist

    Article Title: A lysin motif effector subverts chitin‐triggered immunity to facilitate arbuscular mycorrhizal symbiosis

    doi: 10.1111/nph.16245

    Figure Lengend Snippet: RiSLM ) is highly induced in intraradical mycelium (IRM) in a wide range of host plants. (a) RiSLM is highly induced in five hosts Medicago truncatula (Medicago), Allium schoenoprasum (Chives), Nicotiana benthamiana (Nicotiana), Solanum lycopersicum (Tomato) and Lotus japonicus (Lotus) compared with germinating spores (GS). Transcripts per million (TPM) was used to represent expression levels. Error bars represent SE from either three replicates (for Medicago, Chives, and Nicotiana) or two replicates (for Lotus and Tomato), collected from independent RNA sequencing (RNAseq) experiments. (b) Quantitative PCR analysis, showing that RiSLM (relative to Rhizophagus irregularis elongation factor 1α , RiEF ) is induced during arbuscular mychorrizal colonization and arbuscule (ARB) development as represented by relative expression of RiEF and the ARB‐specific phosphate transporter ( MtPT4 ) normalized by Medicago elongation factor 1α ( MtEF ). Error bars represent SE from three biological replicates. (c) RiSLM expression based on RNAseq analyses of laser microdissected ARBs and IRM compared with extraradical mycelium (ERM) and GS. Error bars represent SE from three biological replicates. For all figures, different characters indicate significant differences (least significant difference P

    Article Snippet: RNA isolation and quantitative PCR For all experiments, total RNAs were isolated using a Qiagen plant RNA mini kit or Omega EZNA Plant RNA kit (Omega Bio‐Tek, Norcross, GA, USA) following the manufacturer’s instructions, including on‐column DNAse treatment.

    Techniques: Expressing, RNA Sequencing Assay, Real-time Polymerase Chain Reaction

    Host‐induced gene silencing of RiSLM ) reduces mycorrhization in Medicago truncatula . (a) Frequency (F%) remains the same, whereas mycorrhization intensity in the root (M%) and arbuscule abundance in the root (A%) are reduced in RiSLM ‐silenced roots. For the empty vector control (EV), 12 biological replicates were used. For RiSLM RNA interference (RNAi), 10 biological replicates were used. (b) Quantitative PCR analysis of control and RNAi roots showing RiSLM expression level relative to Rhizophagus irregularis elongation factor 1α ( RiEF ) and MtPT4 expression levels relative to Medicago elongation factor 1α ( MtEF ). Successful silencing of RiSLM (relative to RiEF ) reduces RiEF and MtPT4 expression (relative to MtEF ), indicating reduced mycorrhization and arbuscule abundance. Twelve replicates (individual transgenic roots) were used for EV. Eleven replicates were used for RNAi . Student’s t ‐test P ‐values are indicated for (a) and (b). For all box plots, boxes represent interquartile range (IQR) and whiskers represent 1.5IQR. (c, d) Wheat germ agglutinin‐Alexa Fluor 488 staining of mycorrhization in (d) RiSLM ‐silenced roots and (c) control roots (c). Bars, 200 µm.

    Journal: The New Phytologist

    Article Title: A lysin motif effector subverts chitin‐triggered immunity to facilitate arbuscular mycorrhizal symbiosis

    doi: 10.1111/nph.16245

    Figure Lengend Snippet: Host‐induced gene silencing of RiSLM ) reduces mycorrhization in Medicago truncatula . (a) Frequency (F%) remains the same, whereas mycorrhization intensity in the root (M%) and arbuscule abundance in the root (A%) are reduced in RiSLM ‐silenced roots. For the empty vector control (EV), 12 biological replicates were used. For RiSLM RNA interference (RNAi), 10 biological replicates were used. (b) Quantitative PCR analysis of control and RNAi roots showing RiSLM expression level relative to Rhizophagus irregularis elongation factor 1α ( RiEF ) and MtPT4 expression levels relative to Medicago elongation factor 1α ( MtEF ). Successful silencing of RiSLM (relative to RiEF ) reduces RiEF and MtPT4 expression (relative to MtEF ), indicating reduced mycorrhization and arbuscule abundance. Twelve replicates (individual transgenic roots) were used for EV. Eleven replicates were used for RNAi . Student’s t ‐test P ‐values are indicated for (a) and (b). For all box plots, boxes represent interquartile range (IQR) and whiskers represent 1.5IQR. (c, d) Wheat germ agglutinin‐Alexa Fluor 488 staining of mycorrhization in (d) RiSLM ‐silenced roots and (c) control roots (c). Bars, 200 µm.

    Article Snippet: RNA isolation and quantitative PCR For all experiments, total RNAs were isolated using a Qiagen plant RNA mini kit or Omega EZNA Plant RNA kit (Omega Bio‐Tek, Norcross, GA, USA) following the manufacturer’s instructions, including on‐column DNAse treatment.

    Techniques: Plasmid Preparation, Real-time Polymerase Chain Reaction, Expressing, Transgenic Assay, Staining