四川大学学报（自然科学版）2017，Vol.54Issue(3)：653-657,5.DOI:10.3969/j.issn.0490-6756.2017.03.038
一种改进的小非编码RNA cDNA文库构建方法
An improved method of sncRNA cDNA library construction
摘要
Abstract
5S and 5.8S rRNA were removed efficiently by RNA Antisense Purification (RAP).More 5'-monophosphorylated RNAs were acquired after using Tobacco Acid Pyrophosphatase (TAP) and RNA 5'sncRNA Polyphosphatase,which resulted in the increasement of the coverage of the cDNA library.To avoid using the adenylated adapter,a poly(A) structure was added to sncRNA 3'end and then reverse transcription was carried out with oligo(dT)16 VN-adapter primers.This improved sncRNA cDNA library contained 3 × 105 independent clones and the recombination rate was 95 ％.The sequencing results showed that the sncRNA cDNA sequences had good integrity by employing this improved and cost-effective method,which laid the foundation for the further study of sncRNA.关键词
sncRNA/cDNA文库/poly(A)/RNA反义链纯化(RAP)Key words
sncRNA/cDNA library/poly(A)/RAP分类
生物科学引用本文复制引用
杜光石,罗发涛,肖燕,陈红军,吴传芳..一种改进的小非编码RNA cDNA文库构建方法[J].四川大学学报（自然科学版）,2017,54(3):653-657,5.基金项目
国家自然科学基金项目(31371325) （31371325）
