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An Efficient Method for the Isolation of Highly Purified RNA from Seeds for Use in Quantitative Transcriptome Analysis
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Biochemistry
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JoVE Journal Biochemistry
An Efficient Method for the Isolation of Highly Purified RNA from Seeds for Use in Quantitative Transcriptome Analysis

An Efficient Method for the Isolation of Highly Purified RNA from Seeds for Use in Quantitative Transcriptome Analysis

DOI:
10.3791/55008-v

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06:31 min

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January 11, 2017

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, ,
1Laboratory of Biological Diversity, Department of Evolutionary Biology and Biodiversity,National Institute for Basic Biology, 2Department of Basic Biology,SOKENDAI (The Graduate University for Advanced Studies), 3Department of Cell Biology,National Institute for Basic Biology

Chapters

  • 00:05Title
  • 00:55Extraction of Total RNA from Plant Seeds
  • 03:05Verification of RNA Quality
  • 03:31Reverse Transcription of Total RNA from Seeds
  • 04:23Quantitative Real-time PCR Analysis
  • 05:01Results: Modified RNA Extraction Process Effectively Removes Inhibitory Compounds
  • 06:10Conclusion

Summary

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We have succeeded in establishing a method for RNA isolation from plant seeds containing large amounts of oils, proteins, and polyphenols, which have inhibitory effects on high-purity RNA isolation. Our method is suitable for monitoring the expression of genes with low level transcripts in seeds.

Tags

RNA ExtractionSeedQuantitative Transcriptome AnalysisArabidopsis ThalianaPVPCell Lysis BufferCentrifugationHomogenizationSpin Column-based MethodHigh-purity RNACereal BiologyGene ProfilesTranscripts

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