GSM8290834: MVC, scRNAseq; Mus musculus; RNA-Seq
Whole live muscle cells were freshly isolated by FACs sorting Sequential crosslinking was performed with the following steps: Whole muscle cells were isolated  with an additional step of viability validation by PI staining. Red blood cells were also eliminated by ACK buffers (150M NH4Cl, 100mM KHCO3, 10mM EDTA-2Na) before sorting. After sorting, live cells were washed with 0.04% BSA in PBS for twice and resuspended in the BSA solution at an appropriate concentration (300-1200 cells/μl). Suspended cells were counted under a microscope and Typan blue was used to examine the cell viability. After the confirmation of cell number and viability, following the 10x manufacturer's instructions for generation of Gel Bead-In Emulsions (GEMs) using the 10X Chromium system. Next GEMs were proceeded with in-GEM reverse transcription (RT) under manufacturer's instructions. After RT, two steps of DNA cleaning up and purification were performed to obtain the purified cDNA sample for further library construction with the manufacturer's protocols. Bioanalyzer analysis and qPCR were used to measure the quality of DNA libraries including the DNA size and purity. Finally, DNA libraries were sequenced on the Illumina Illumina HiSeq X ten platform.