GSM6886900: biol rep 3- pancreatic Trip12 knockout acinar cells (KO_787); Mus musculus; RNA-Seq
Total RNA were isolated using Trizol reagent (Life Technologies) following manufacturer's recommendations. Preliminary quality control of total mRNA was ensured by agarose gel electrophoresis and NanoDrop measurement. Sample quantitation and integrity were performed by Nanodrop and Agilent 2100, respectively. mRNA libraries were constructed using poly A enrichment preparation. Messenger RNA was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers followed by the second strand cDNA synthesis. The library was ready after end repair, A-tailing, adapter ligation, size selection, amplification, and purification. The library was checked with Qubit and real-time PCR for quantification and bioanalyzer for size distribution detection. Quantified libraries will be pooled and sequenced on Illumina platforms, according to effective library concentration and data amount