GSM2898275: 2h PH rep2; Mus musculus; RNA-Seq
The liver were removed, flash frozen in liquid nitrogen and then were stored in -70℃. Total RNA were prepared from the liver samples using TRIZOL reagent (Invitrogen), and three RNA samples from the same time point were pooled. There were two pooled samples in each time point. The total  RNA samples are first treated with DNase I to degrade any possible DNA contamination. Then the mRNA is enriched by using the oligo(dT) magnetic beads (for eukaryotes). Mixed with the fragmentati on buffer, the mRNA is fragmented into short fragments (about 200 bp). Then the first strand of cDNA is synthesized by using random hexamer-primer. Buffer, dNTPs, RNase H and DNA polymerase I are added to synthesize the second strand. The double strand cDNA is purified with magnetic beads. End reparation and 3' -end single nucleoti de A (adenine) addition is then performed. Finally, sequencing adaptors are ligated to the fragments. The fragments are enriched by PCR amplification. During the QC step, Agilent 2100 Bi oanaylzer and ABI StepOnePl us Real-Time PCR System are used to qualify and quantify of the sample library.
GEO Accession
GSM2898275