GSM1873345: L-Input-1; Homo sapiens; RIP-Seq
Cells were crosslinked with 0.3% formaldehyde for 10 min at room temperature and lysed with RIPA lysis buffer for 10 min on ice before sonication. The supernatant was collected and incubated with RIP antibodies for 4 to 6 hours. After washing with RIPA buffer, RNA was eluted from the beads with 100 µl NaHCO3 and 1% SDS in the presence of proteinase K and RNase inhibitor at room temperature for 10 min with occasional vortex. The eluted material was de-crosslinked at 65°C for 45 – 60 min before purification of RNA using Trizol LS reagent (Life Technology). DNase I treatment was performed to remove any residual DNA. RNA-seq libraries were constructed with TruSeq Stranded mRNA Library Prep Kit from Illumina according to the manufacturer’s manual.
GEO Accession
GSM1873345