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单次读取和配对的最终从10微克总RNA的mRNA - Seq的Illumina的图书馆

JoVE Journal
Biology

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JoVE Journal Biology

Single Read and Paired End mRNA-Seq Illumina Libraries from 10 Nanograms Total RNA

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单次读取和配对的最终从10微克总RNA的mRNA - Seq的Illumina的图书馆

DOI:

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14:49 min

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October 27, 2011

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Srikumar Sengupta, Jennifer M. Bolin, Victor Ruotti, Bao Kim Nguyen, James A. Thomson^2,3, Angela L. Elwell, Ron Stewart

¹Regenerative Biology,Morgridge Institute for Research, ²Department of Cell & Regenerative Biology,University of Wisconsin, ³Department of Molecular, Cellular, & Regenerative Biology,University of California

Chapters

00:05Title
02:02Procedural Flow Chart
03:19cDNA Synthesis
06:31End Repair, Paired End Library
07:37Adding ‘A’ Bases to the 3′ End of the DNA Fragments
08:31Adapter Ligation
10:32Size Selection/Gel Purification
11:20Evaluation of Single-Read and Paired-End Sequencing Libraries
14:18Conclusion

Summary

Automatic Translation

在这里，我们描述了读取和配对的单端Illumina公司的mRNA - Seq的测序基础上的T7线性RNA扩增的基因表达分析库的制备方法。该协议只需要10微克启动总RNA，并产生了高度一致库代表整个笔录。

Tags

MRNA-Seq Illumina Libraries Global Gene Expression Mutation Analysis Allele-specific Expression Genome-wide Analyses Non-sequenced Genomes Transcript Copy Numbers Illumina’s Genome Analyzer Paired End Sequencing Alternate Splice Junctions Insertions And Deletions De Novo Transcriptome Assembly Limited Starting Material Nanograms Of Total RNA PCR Amplification Minimal PCR Amplification

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