GSM2824213: DHX36-iso2 overexpression 2; Homo sapiens; RIP-Seq
Cells were lysed in NP-40 lysis buffer and lysate was clarified by centrifugation. First RNase T1 was followed by immunoprecipitation with FLAG-M2 antibody After washing, second RNase T1 digestion was performed, 3´ends of the RNA fragments were dephosphorylated using CIP, and RNA was radioactively 5´end-labeled using l T4 PNK and 32P-γ-ATP. RNA-protein complexes were separated by SDS-PAGE and excised. After proteinase K treatment, small RNA cDNA library preparation was performed. Preadenylated, barcoded 3´adapter oligonucleotide was ligated followed by ligation of a 5´adapter. Ligation products were reverse transcribed and amplified by PCR.
GEO Accession
GSM2824213