mRNA fraction of TIG-1 cells (PDL=52)
mRNAs in 400 cells of TIG-1 cells with population doubling levels (PDL) = 52 were reverse-transcribed with template-switch oligos and anchored-poly(dT)28 primers, flanking 8-bp UMI, 10-bp cell barcode, TruSeq read 1 sequence, and PCR handle. After digesting residual primers by exonuclease I, the full-length cDNA was amplified by 15 cycles of PCR, and purified by 0.7x AMPure XP (Beckman Coulter), pooling 16 samples. The purified cDNA samples of 800 pg were used for constructing a sequencing library (Nextera XT DNA Library Preparation Kit, Illumina) using Illumina TruSeq P5 and Nextera P7 adaptors to enrich the reads near the 3 prime regions of mRNA, and the purified by AMPure XP.