GSM8325446: 170308-07, NPDR; Homo sapiens; miRNA-Seq
Participants were instructed to fast for 8-12 hours, and approximately 5ml of venous blood was collected using vacuum tubes containing citrate anticoagulant at room temperature in the early morning. The whole blood was centrifuged at 1000g for 10 minutes, and the plasma was collected and stored at -80°C for further use. Total RNA of each sample was used to prepare the miRNA sequencing library, which included the following steps:  1) 3'-adaptor ligation;  2) 5'-adaptor ligation;  3) cDNA synthesis;  4) PCR amplification;  5) size selection of ~135-155 bp PCR amplified fragments (corresponding to ~15-35 nt small RNAs).  The libraries were denatured as single-stranded DNA molecules, captured on Illumina flow cells, amplified in situ as clusters and finally sequenced for 50 cycles on Illumina NextSeq 500 per the manufacturer's instructions.