3-Adp_run1
First, a 5'-adenylated RNA adapter was ligated to the 3'-end of the sRNA sample. Then, 5'-PPPstructures were removed using tobacco acid pyrophosphatase . This was followed by ligating a secondRNA adapter to the 5'-monophosphate of the RNA. First-strand cDNA synthesis was primed using the3' adapter primer and was carried out with M-MLV reverse transcriptase. The resulting cDNA wasPCR-amplified to about 20-30 ng/l using a high fidelity DNA polymerase. PCR cycles performed andthe barcode sequence, which is part of the 3'-TruSeq sequencing adapter, is included in Table 2. ThecDNA was purified using the Agencourt AMPure XP kit (Beckman Coulter Genomics). TrueSeq_Sense_primer: 5'-AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCT-3'TrueSeq_Antisense_primer: 5'-CAAGCAGAAGACGGCATACGAGAT-NNNNNN-GTGACTGGAGTTCAGACGTGTGCTCTTCCGATC-3'
SUB14703285
3-Adp_run1