直接测序法检测RNA 编辑

Mehdi Jabnoune; David Secco; Cécile Lecampion; Christophe Robaglia; Qingyao Shu; Yves Poirier

doi:10.21769/BioProtoc.1410

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Peer-reviewed

RNA Editing Detection by Direct Sequencing

直接测序法检测RNA 编辑

Mehdi Jabnoune email

DS David Secco

CL Cécile Lecampion

CR Christophe Robaglia

QS Qingyao Shu

YP Yves Poirier

发布: 2015年03月05日第5卷第5期 DOI: 10.21769/BioProtoc.1410 浏览次数: 8822

评审: Ru ZhangAnonymous reviewer(s)

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The authors used this protocol in:

Cover of The Plant Cell, featuring study using the protocol.

Oct 2013

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Abstract

RNA editing is a widespread post-transcriptional phenomenon through which primary RNA sequences are altered by nucleotide insertion/deletion or base conversion. It occurs in a variety of organisms and cooperates with alternative splicing in increasing both proteomic and transcriptomic complexity. We describe here a method allowing RNA editing events detection by performing direct sequencing of both genomic DNA and cDNA from the same source.

Materials and Reagents

DNeasy Plant mini kit (QIAGEN, catalog number: 69104 )
RNeasy mini kit (QIAGEN, catalog number: 74104 )
pJET PCR cloning system (Thermo Fisher Scientific, Fermentas, catalog number: K1231 )
Recombinant DNase I (Life Technologies, Ambion^®, catalog number: AM2235 )
M-MLV reverse transcriptase (Promega corporation, catalog number: 28025-013 )
Oligo(dT) 15 Primer (500 mg) (Promega corporation, catalog number: C1101 )
KOD high fidelity polymerase (TOYOBO, catalog number: KOD-201 200U )
Zymoclean Gel DNA Recovery Kit (ZYMO RESEARC, catalog number: D4001 )

Equipment

NanoDrop (Thermo Fisher Scientific)

Procedure

English

中文翻译

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如何引用

Jabnoune, M., Secco, D., Lecampion, C., Robaglia, C., Shu, Q. and Poirier, Y. (2015). RNA Editing Detection by Direct Sequencing. Bio-protocol 5(5): e1410. DOI: 10.21769/BioProtoc.1410.