RNA Editing Detection by Direct Sequencing
直接测序法检测RNA 编辑
发布: 2015年03月05日第5卷第5期 DOI: 10.21769/BioProtoc.1410 浏览次数: 9632
评审: Ru ZhangAnonymous reviewer(s)
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参见作者原研究论文
The authors used this protocol in:
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
文章信息
版权信息
© 2015 The Authors; exclusive licensee Bio-protocol LLC.
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.
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分类
植物科学 > 植物分子生物学 > RNA > RNA 测序
分子生物学 > RNA > RNA 测序
分子生物学 > RNA > RNA 检测
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