GSM5680799: miRNA In labor myometrium rep2; Homo sapiens; miRNA-Seq
Myometrium were removed, flash frozen on dry ice, and RNA was harvested using Trizol reagent. Total RNA was separated by polyacrylamide gel electrophoresis (PAGE) and small RNA regions corresponding to the 18–30 nt bands in the marker lane (14-30 ssRNA Ladder Marker, TAKARA) were excised and recovered. Then the small RNAs were ligated to adenylated 3' adapters annealed to unique molecular identifiers (UMI), followed by the ligation of 5' adapters. The adapter-ligated small RNAs were transcribed into cDNA, and subsequently enriched by PCR. The target fragments 110~130 bp were selected by agarose gel electrophoresis, and purified by QIAquick Gel Extraction Kit (QIAGEN, V alencia, CA). The library was quality and quantitated in two methods: check the distribution of the fragments size using the Agilent 2100 bioanalyzer, and quantify the library using real-time quantitative PCR (qPCR) (TaqMan Probe). The final ligation PCR products were sequenced using the BGISEQ-500 platform (BGI-Shenzhen, China).
GEO Accession
GSM5680799