title: GSM6670680: EM_22_04; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA were isolated from tissues by using the Trizol Reagent according to the manufacture's protocol The poly-A containing mRNA molecules were purified from the 3 μg of total RNA by using poly-T oligo-attached magnetic beads. The cleaved RNA fragments were reversely transcribed into first strand cDNA using random hexamers, following by second strand cDNA synthesis using DNA Polymerase I and RNase H. The cDNA fragments were purified, end blunted, 'A' tailed, and adaptor ligated. PCR was used to selectively enrich those DNA fragments that have adapter molecules on both ends and to amplify the amount of DNA in the library. The number of PCR cycles was minimized to avoid skewing the representation of the library. The library were qualified by Agilent 2100 bioanalyzer and quantified by Qubit and qPCR.