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A non-radioactive, improved PAR-CLIP and small RNA cDNA library preparation protocol


ABSTRACT: Streamlined protocol for fluorescence-based PAR-CLIP (fPAR-CLIP) that eliminates the need to use radioactivity. Protocol is based on direct ligation of a fluorescently labeled adapter to the 3’end of crosslinked RNA on immobilized ribonucleoproteins, followed by isolation of the RNA and efficient conversion into cDNA without the previously needed size fractionation on denaturing polyacrylamide gels. These improvements cut the experimentation time from 4-5 to 2 days and increases sensitivity by 10-100-fold.

ORGANISM(S): Homo sapiens

PROVIDER: GSE163925 | GEO | 2020/12/29

REPOSITORIES: GEO

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