polyC_run1
The sRNA sample was poly(C)-tailed using poly(A)polymerase (PAP) in the presence of CTP instead of ATP. The reaction was carried our for 2 h at37C, then additional PAP was added and the sample was further incubated at 37C for another twohours. Then, 5'-PPP structures were removed using tobacco acid pyrophosphatase. Afterwards, aRNA adapter was ligated to the 5'-monophosphate of the RNA. First-strand cDNA synthesis wasperformed using an oligo(dT)-adapter primer and M-MLV reverse transcriptase. The resulting cDNAwas PCR-amplified to about 20-30 ng/l using a high fidelity DNA polymerase. PCR cycles performedand the barcode sequence, which is part of the 3'-TruSeq sequencing adapter
SUB14703285
polyC_run1