title: GSM9346983: Isolated injured cardiac tissue from WT mice; Mus musculus; RNA-Seq
library_construction_protocol: 3 days post cryo injury WT and ABCC6 KO mice scar regions of the heart were harvested and digested by 0.1 µg/mL liberase TH (Sigma, 5401151001). After digestion, cells were incubated with 10 µM Calcein AM (Abcam, ab141420) and live cells were collected by flow cytometry Single-cell gene expression libraries were created using Chromium Next GEM Single Cell 3' (v3.1 Chemistry) (10x Genomics), Chromium Next GEM Chip G Single Cell Kit (10x Genomics), and Single Index Kit T Set A (10x Genomics) according to the manufacturer's instructions. Briefly, samples were loaded to target 10,000 cells to form GEMs and barcode individual cells. GEMs were then cleaned cDNA and libraries were also created according to the manufacturer's instructions. Library quality was assessed using the 4200 TapeStation System and D1000 ScreenTape (Agilent) and Qubit 2.0 (Invitrogen) for concentration and size distribution. Samples were sequenced using Novaseq 6000 (Illumina) using 100 cycles. 200 M reads were targeted for each sample, targeting 20,000 reads per cell. Raw reads were processed using the Cell Ranger Pipeline (10x Genomics)