GSM5219400: d6_S1 (scRNA-seq); Mus musculus; RNA-Seq
Single cell RNA libraries were prepared using Chromium Single Cell 3' Library Kit v2 (10X genomics, 120234) and the generated libraries were sequenced using Illumina NextSeq 500 platform. Reprogrammable cells at day 6 or 12 were prepared as instructed in the 10X Genomics cell preparation guidelines. Briefly, cells were trypsinized and centrifuged at 1000 RPM for 3 minutes, then were washed twice with PBSx1 containing 0.04% BSA and cleaned from cell debris and large clumps by filtering throw mesh paper. Next, resuspended cells were subjected to dead cell removal kit (MACS, 130-090-101) to remove any non-viable cells. Cell viability were estimated using trypan blue staining. Cells were then resuspended in PBSx1 with 0.04% BSA at the concentration 1000 cells/µl and 4000 cells from each condition were subjected to 10x Genomics.