GSM4972361: Day 7 post-MI; Mus musculus; RNA-Seq
We sorted total viable CD45+ leukocytes from the mouse heart. Five samples were analyzed: control, after 1, 3, 5, and 7 days of myocardial infarction. The single-cell library preparation relied on a commercially available droplet method, the 10x Genomic Chromium System (10x Genomic Inc., San Francisco, CA), with 10x Genomics Single Cell 3' v3 Reagent kit according to the manufacturer's protocol. In brief, dissociated cells were counted by hemocytometer (ThermoFisher) and 16,000 cells per sample were added to each channel. The cells were then partitioned into gel beads in emulsion (GEMs) in the Chromium instrument, where cell lysis and barcoded reverse transcription of RNA occurred. cDNA was synthesized and amplified for 14 cycles. cDNA clean-up was performed using a SPRIselect Reagent Kit (Beckman Coulter, Brea, CA). 50 ng of the amplified cDNA were used for each sample to construct indexed sequencing libraries.