GSM7774598: H1, 1-10 KO, scRNA; Homo sapiens; RNA-Seq
Routinely cultured hESCs were digested into single cells with TrypLE (Thermo Fisher) according to the manufacturer's protocol. Then cells were re-suspended in 1 x DPBS (Thermo Fisher) and stained with a Viability Dye. High viability samples were subject to scRNA-seq library preparation. Libraries were prepared using a method established in our lab which is adapted from Microwell-seq. Briefly, hESCs and barcoded beads were first distributed into wells of a microwell plate, with each well containing a single cell and a single bead. Then, polyA RNA from the cell lysate were captured and retrotranscripted into cDNA. The pooled cDNA were pre-amplified by PCR, then fragmented and tagmented with Tn5 Transposase. Finally, Illumina p5 and p7 sequences and sample indexes were introduced by another round of PCR. Fragment size of 300-600 bp were selected for sequencing.