Lysates were clarified from sonicated Huh7 cells and PCBP2-RNA complexes were isolated with an anti-PCBP2 antibody. Libraries were prepared according to the standard iCLIP protocol (PMID: 24184352), substituting the 3' RNA adaptor for one with a 3'-Biotin blocking moity. Subsequent steps utalized a Streptavidin-Biotin capture strategy to eliminate the need for precipiations between biochemical reactions. Each sequencing read conatins a 13 nt random (D's and N's) and experimental (XXXX) "DDDNNXXXXNNNN". The experimental barcodes are described in (PMID: 24184352)
GEO Accession
GSM1447363