GSM8459880: Ctrl2 [RNA-seq]; Bos taurus; RNA-Seq
The RNA-seq libraries were constructed by E-GENE Co. Ltd as follows. Briefly, the total RNA of each sample was extracted using Invitrogen TRIzol® Reagent and then treated with RNase-free DNase I for30 min according to the manufacturer's protocols. The poly(A) containing mRNA were purified using Oligo(dT) Beads from about 1 ug total RNA. The captured mRNA was firstly fragmented into 100-200nt using divalent cations at elevated temperature. The fragmented mRNA was reverse transcripted with SuperScript II and then converted to double-stranded cDNA using RNaseH and DNA Pol I by random priming. After purification, the double-stranded cDNA was subjected to blunt-ending, dA addition to 3'-end and adapter ligation. Finally, PCR was carried out to enrich the adapter ligated cDNA and the libraries were analyzed by Agilent Bioanalyzer 2100 and quantified by qPCR before sequenced by the Illumina sequencing platform.