GSM8128525: C2C12 myoblasts, LMNA-R482L, rep1; Mus musculus; RNA-Seq
After C2C12 cells culturing in 6-well plates in proliferation medium (DMEM supplemented with 4.5 g/L D-glucose, L-glutamine, penicillin-streptomycin, and 20% FCS), C2C12 cells were harvested using Trypsin, washed with PBS and resuspended in PBS. Library was performed according to the manufacturer's instructions (Single Cell 3' v3.1 protocol, 10X Genomics). Briefly, C2C12 cells suspensions with the targeted cell recovery rate 5000 cells per sample were loaded onto 10x Genomics Single Cell fluidics chips to generate GEMs (Gel Beads-in-emulsion) which include gel beads coated with specific oligonucleotide barcodes to index each cell's transcriptome and Unique Molecular Identifiers (UMI) to define individual transcripts. Uniquely barcoded RNAs were reverse transcribed, and individual cDNAs were amplified using a bench top Veriti™ 96-well thermal cycler (Applied Biosystems). Each library was barcoded with different indexes from a Chromium i7 Multiplex Kit. Barcoded libraries were then pooled and sequenced on the Illumina NextSeq 2000 system.