GSM6503383: Runx1, replicate 2, RIP-seq; Mus musculus; RIP-Seq
10 μL Protein A Dynabeads were added to 500 μL RIPA buffer containing proteinase inhibitor cocktail, rotated 1-2 min at RT and placed on the magnet stand to remove supernatant. Then pre-treated beads were resuspended with 50 μL RIPA buffer containing 1 μg indicated RIP antibodies, rotated and incubated at 4 °C for 4-6 hours, and then placed on the magnet stand to remove the supernatant. The cDNA libraries were generated as previously reported with a litter modification[16]. Instead of a single cell, a certain volume of RNA solution was added into the reverse transcription system for reverse transcription After that, cDNA was sonicated to ~300 bp fragments by Covaris S2 system and purified with QIAGEN PCR purification kit. Then libraries were generated using KAPA Hyper Prep Kit Illumina® platforms following the manufacture's protocol.