期刊
NUCLEIC ACIDS RESEARCH
卷 50, 期 4, 页码 -出版社
OXFORD UNIV PRESS
DOI: 10.1093/nar/gkab1158
关键词
-
资金
- IMMAGINA BioTechnology S.r.l. internal funding
- AFMTelethon [22129]
- Caritro Foundation
We developed a PCR-free library preparation method called circAID-p-seq for selective 3' phospho-RNA sequencing. By applying it to ribosome profiling, we were able to accurately and quickly sequence phospho-RNA fragments from eukaryotic cells and tissues, and depict ribosome occupancy in transcripts.
Most RNA footprinting approaches that require ribonuclease cleavage generate RNA fragments bearing a phosphate or cyclic phosphate group at their 3 ' end. Unfortunately, current library preparation protocols rely only on a 3 ' hydroxyl group for adaptor ligation or poly-A tailing. Here, we developed circAID-p-seq, a PCR-free library preparation for selective 3 ' phospho-RNA sequencing. As a proof of concept, we applied circAID-p-seq to ribosome profiling, which is based on sequencing of RNA fragments protected by ribosomes after endonuclease digestion. CircAID-p-seq, combined with the dedicated computational pipeline circAidMe, facilitates accurate, fast and highly efficient sequencing of phospho-RNA fragments from eukaryotic cells and tissues. We used circAID-p-seq to portray ribosome occupancy in transcripts, providing a versatile and PCR-free strategy to possibly unravel any endogenous 3 '-phospho RNA molecules.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据