RTL-P: a sensitive approach for detecting sites of 2′-O-methylation in RNA molecules
文献类型：期刊论文
| 作者 | Dong, Zhi-Wei; Shao, Peng; Diao, Li-Ting; Zhou, Hui; Yu, Chun-Hong; Qu, Liang-Hu |
| 刊名 | Nucleic acids research
 |
| 出版日期 | 2012 |
| 英文摘要 | 2'-O-methylation is present within various cellular RNAs and is essential to RNA biogenesis and functionality. Several methods have been developed for the identification and localization of 2'-O-methylated sites in RNAs; however, the detection of RNA modifications, especially in low-abundance RNAs and small non-coding RNAs with a 2'-O-methylation at the 3'-end, remains a difficult task. Here, we introduce a new method to detect 2'-O-methylated sites in diverseRNA species, referred to as RTL-P [Reverse Transcription at Low deoxy-ribonucleoside triphosphate (dNTP) concentrations followed by polymerase chain reaction (PCR)] that demonstrates precise mapping and superior sensitivity compared with previous techniques. The main procedures of RTL-P include a site-specific primer extension by reverse transcriptase at a low dNTP concentration and a semi-quantitative PCR amplification step. No radiolabeled or fluorescent primers are required. By designing specific RT primers, we used RTL-P to detect both previously identified and novel 2'-O-methylated sites in human and yeast ribosomal RNAs (rRNAs), as well as mouse piwi-interacting RNAs (piRNAs). These results demonstrate the powerful application of RTL-P for the systematic analysis of fully or partially methylated residues in diverse RNA species, including low-abundance RNAs or small non-coding RNAs such as piRNAs and microRNAs (miRNAs). |
| 收录类别 | SCI |
| 原文出处 | http://nar.oxfordjournals.org/content/40/20/e157.full.pdf+html |
| 语种 | 英语 |
| 源URL | [http://ir.siat.ac.cn:8080/handle/172644/4267]  |
| 专题 | 深圳先进技术研究院_医药所 |
| 作者单位 | Nucleic acids research |
| 推荐引用方式 GB/T 7714 | Dong, Zhi-Wei,Shao, Peng,Diao, Li-Ting,et al. RTL-P: a sensitive approach for detecting sites of 2′-O-methylation in RNA molecules[J]. Nucleic acids research,2012. |
| APA | Dong, Zhi-Wei,Shao, Peng,Diao, Li-Ting,Zhou, Hui,Yu, Chun-Hong,&Qu, Liang-Hu.(2012).RTL-P: a sensitive approach for detecting sites of 2′-O-methylation in RNA molecules.Nucleic acids research. |
| MLA | Dong, Zhi-Wei,et al."RTL-P: a sensitive approach for detecting sites of 2′-O-methylation in RNA molecules".Nucleic acids research (2012). |
入库方式： OAI收割
来源：深圳先进技术研究院
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