GSM7123505: mouse 4254, Ncoa5+/- [m4254_CIA_WT]; Mus musculus; RNA-Seq
A piece of liver was cut from samples collected from mice shortly after euthanasia and freshly frozen in liquid nitrogen. Livers were homogenized in TRIzol and total RNA was extracted using a combined TRIzol and RNeasy Mini Kit (Qiagen) method. RNA quality control was performed using the Nanodrop Spectrophotometer (Thermo Fisher) and Bioanalyzer (Agilent). 250~300bp insert cDNA library preparation was carried out by Novogene. Generally mRNA was enriched using oligo(dT) beads. mRNA is fragmented randomly by adding fragmentation buffer, then the cDNA is synthesized by using mRNA template and random hexamers primer, after which a custom second-strand synthesis buffer (Illumina), dNTPs, RNase H and DNA polymerase I are added to initiate the second-strand synthesis. Second, after a series of terminal repair, A ligation and sequencing adaptor ligation, the double-stranded cDNA library is completed through size selection and PCR enrichment. (Method provided by Novogene)