title: GSM8372222: human_CD8_D12; Homo sapiens; miRNA-Seq
library_construction_protocol: Total RNA was isolated using the TRIzol reagent (Life Technologies) following the manufacturer's instructions. A total of 1 μg of RNA was used for library preparation. First, the 3' SR Adaptor for Illumina was ligated to the small RNA using a 3' Ligation Enzyme. To prevent adaptor-dimer formation, excess 3' SR Adaptor was hybridized with the SR RT Primer for Illumina. Next, the 5' SR Adaptor for Illumina was ligated to the small RNA using a 5' Ligation Enzyme, and first-strand cDNA was synthesized using ProtoScript II Reverse Transcriptase. Each sample was then amplified by PCR using P5 and P7 primers. The PCR products were purified using DNA clean beads, and the purified products ranging from 140-160 bp were recovered and cleaned up using PAGE.