GSM7508029: Liver_Cisplatin_5; Mus musculus; RNA-Seq
Total RNA was isolated from freshly frozen liver tissues from the mice treated with indicated chemotherapy agents by TRIzol reagent (#T9424, Invitrogen). RNA library for RNA-seq was prepared as following: mRNA was purified from total RNA using polyT and then fragmented into 300~350 bp fragments. The first strand cDNA was reverse-transcribed using fragmented RNA and dNTPs (dATP, dTTP, dCTP and dGTP) and second strand cDNA synthesis was subsequently performed. Remaining overhangs of double-strand cDNA were converted into blunt ends via exonuclease/polymerase activities. After adenylation of 3' ends of DNA fragments, sequencing adaptors were ligated to the cDNA and the library fragments were purified. The template was enriched by PCR, and the PCR product was purified to obtain the final library. After library construction, the concentration of library was measured by the Qubit fluorometer. The accurate concentration of cDNA library was again examined using qPCR. The size distribution of library was detected by agarose gel electrophoresis.