GSM2414879: MEL4; Rattus norvegicus; RNA-Seq
Total RNA was isolated using a GenElute Mammalian Total RNA Miniprep kit (Sigma) according to the manufacturer’s instructions. DNase I digestion (On-Column DNase I digestion set, Sigma) was performed using a modified manufacturer’s protocol (15 minutes at 37°C on column). RNA concentrations were determined by spectrophotometry at 260 nm using Nanodrop (ThermoFisher). Moreover, the RNA quality was tested by Agilent 2100 Bioanalyzer (Agilent Technologies). RNA integrity number (RIN) range was 9.0-9.7. Library construction by BGI International; mRNA was enriched by oligo (dT) magnetic beads and fragmented. Then the first strand of cDNA was synthesized by using random hexamer-primer. Buffer, dNTPs, RNase H and DNA polymerase I were added to synthesize the second strand. The double-stranded cDNA was purified with magnetic beads. End reparation, 3’-end single nucleotide A (adenine) addition and sequencing adaptor ligation was then performed. The fragments were enriched by PCR amplification
GEO Accession
GSM2414879