GSM5820435: CNTRL E11.5; Mus musculus; RNA-Seq
Embryos (E9.5 and E11.5) were removed from yolk sac, whole hearts microdissected in diethylpyrocarbonate-treated PBS, dissociated into single cell suspension using Collagenase II (Worthington) and TryPLE. >85-90% viable cells were used to prepare single GEMs using 10XGenomics Chromium Controller. The quality of cDNA were assessed using High Sensitivity  D5000 ScreenTape® on Agilent 2200 TapeStation. After adjusting all four samples (CNTRL vs. matHG-exposed E9.5 and E11.5) to similar concentrations, each single-cell cDNA sample was used for library preparation using Chromium Single Cell 3′ Library & Gel Bead Kit v2 and Chromium i7 Multiplex Kit (10X Genomics,PN-120237, PN-120262) and quantified using High Sensitivity D1000 ScreenTape®. Single-cell libraries were sequenced on the Illumina Hiseq4000 platform with 2 X 150 bp read length using standard Illumina protocol. Chromium Single Cell 3′ Library & Gel Bead Kit v2 and Chromium i7 Multiplex Kit (10X Genomics,PN-120237, PN-120262) Single-cell RNA-seq libraries were sequenced on the Illumina Hiseq4000 platform (2x150 bp read length)