title: RNA-Seq of Homo sapiens: adult female cervix uterus
design: We extract RNA from tissue using TRIzol reagent. The concentration and purity of RNA were measured with a NanoDrop ND-1000 spectrophotometer. We used Bioanalyzer 2100 and agarose electrophoresis to determine whether the RNA was degraded. Downstream experiments required a total amount of RNA greater than 1 g. The first cDNA strand was synthesized by Invitrogen SuperScriptII reverse transcriptase. The second cDNA strand was synthesized from E. coli DNA polymerase I  and RNase H. Then, dUTP solution  was added for cDNA end repair. After the A-tails addition, magnetic bead screening and purification, the second strand was digested with UDG enzyme and a 300bp50bp cDNA library was constructed by PCR. The cDNA library was double-ended PE150 sequenced on the illumina Novaseq6000 platform.