GSM2810845: Mock CLIP-seq exp2; Homo sapiens; RIP-Seq
Cells were lysed with passive lysis buffer. The protocol was adapted from Moore et al., 2014 Anti-T7 antibody was coupled to 50 μl Dynabeads per sample. Following immunoprecipitation, the RNA was dephosphorylated, the 3' linker was ligated and radiolabeled and the 5' linker was ligated on the beads. The 3' linker reaction was performed in a total of 80 μl, with 1x PNK buffer, 1 μM 3' adapter, 800 U T4 RNA ligase 2 truncated K227Q, 80 U RNase out, and 10% PEG 8000. The 5' linker reaction occurred in a total of 80 μl, with 1x PNK buffer, 1 mM ATP, 12.5 nM 5' linker, 80 U RNase OUT and 40 U T4 RNA ligase. The RNA was reverse transcribed and amplified by PCR using LA Takara Taq.
GEO Accession
GSM2810845