title: GSM8759901: CD8 T, Biotin-, rep3; Mus musculus; RNA-Seq
library_name: GSM8759901
library_construction_protocol: The sorted cells were lysed in 50-100 μL TRIzon reagent (CWBIO, CW0580S) by pipetting or vortexing. To the lysate, 20 μL of CHCl3 was added, mixed thoroughly, and the mixture was centrifuged at 12,000 rpm, 4 °C for 15 minutes to transfer the upper clear aqueous phase to a new RNase-free tube. Total RNA was extracted using RNA clean & concentrator kits (Zymo, R1015). mRNA 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. After library quality control, quantified libraries will be pooled and sequenced on Illumina platforms, according to effective library concentration and data amount.