GSM5233839: 10_S3: TF-10 HaCaT; Homo sapiens; RNA-Seq
Trypsinized single cells were washed in PBS (without ca++ or Mg++) with 0.04% BSA and tested for >90% viability. Single cell analysis was performed on a 10xGenomics platform according to 3' v3.1 NextGem Dual Index manual. The 3'-biased cDNA libraries were constructed through the following steps: cDNA fragmentation, end repair & A-tailing, and size selection by SPRIselect beads, adaptor ligation, and sample index PCR amplification, and then SPRIselect beads size selection again. The final constructed 3'-biased single cell libraries were sequenced by Illumina Nextseq500 machine, targeting total reads per cell for 20,000 at minimum, and the sequencing cycles consisted of 28bp for read 1, 90 bp for read 2, and 10 bp for i7 index, and 10bp for i5 index.