GSM5939218: SCS02; Mus musculus; RNA-Seq
Total RNA was isolated from the bulk tissue using the RNeasy® Micro Kit (Qiagen, #74004) according to manufacturer's instructions. 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. Then the quantified libraries were pooled and sequenced for 150pb paired-read and performed on an Illumina Novaseq6000 platform using 2x150 sequencing to read depth of > 40 million reads per sample at Novogene Co. Ltd. genomics core facility (Cambridge, UK).