GSM9629038: RNAseq_WT_rep3; Mus musculus; RNA-Seq
5% of Drosophila S2 cells was added to the collected CH12F3 cells as the spike-in control. Total RNA was extracted by Trizol reagent (thermo fisher 15596018CN). We used 1-2ug total RNA to construct RNA-seq library. Genomic DNA was firstly removed, RNA was reverse-trascripted by using oligo-dT primer sets to obatin cDNA.cDNA was tagmentated and then purified before PCR. We used High-Fidelity 2X PCR Master Mix (NEB, M0541) and illumina index primer sets to construct libraries. PCR was set up as follows: 72 °C, 3 min;95 °C, 3 min; [98 °C, 20 s, 60 °C, 15 s, 72 °C, 30 s] × 11-20 cycles; 72 °C, 5 min,4 °C hold. The amplified libraries were purified by AmpureXP beads and were sequenced via paired end 150bp sequencing on a NovaSeq 6000 (Illumina) sequencer.