GSM1572155: HeLa transfected with a control siRNA; Homo sapiens; miRNA-Seq
Total RNAs were extracted by TRIzol (Invitrogen) following the manufacturer's instructions. To prepare pre-miRNA cDNA libraries, total RNAs were separated on 15% urea-PAGE and RNAs of 40-120nt were gel-purified. Size fractionated RNAs were ligated to 3ʹ adapter by using T4 RNA ligase 2, truncated (NEB). The 3ʹ adapter-ligated RNA was separated on 12.5% urea-PAGE and RNAs of 60-140nt were gel-purified. Size fractionated RNAs were hybridized with an RT primer that is complementary to the 3ʹ adapter and reverse-transcribed by using SuperScript III (Life Technologies) and the cDNA was PCR-amplified with miRNA specific primers using Phusion DNA polymerase (NEB). The cDNA libraries were separated on 6% native polyacrylamide gel and RNAs of 150-225nt.
GEO
GSM1572155