GSM8015322
Cells were lysed on ice using 50 μl wash buffer for 10 min. Next, 1 μl SUPERase·In RNase inhibitor (Ambion, AM2696) and 4 μl RQ1 DNase (Promega, M6101) were applied to the lysate and incubated at 37 °C for 3 min. RNA immunoprecipitation of the samples was performed. The immunoprecipitated RNAs were then fragmented by MNase and dephosphorylated. Then a series of steps were performed to include, reverse transcription, first-strand cDNA capture by streptavidin beads, poly(A) tailing, pre-PCR, IVT, RNA purification, RT, PCR barcoding and deep sequencing.