GSM8546133: ITA, woman 4, quercetin; Homo sapiens; RNA-Seq
Thoracic arteries were harvested in operating room and snap frozen within 15min and stored at -80oC. Frozen segments were processed using GentleMACS Octo Dissociator and its nuclei lysis buffer, according the the manufacturer's instructions. Nuclei were then washed, filtered (70µm and 20µm) and resuspended in PBS-BSA 0.1%. Libraries were prepared using the Next GEM Single Cell 3' Dual Indexing kit (10X Genomics) using a Chromium Controller instrument to generate single-nuclei gel bead-in-emulsion (GEMs). Then, GEMs were reverse transcribed into cDNAs that were then amplified and cleaned up using SPRIselect Reagent Kit (Beckman Coulter). The barcoded sequencing libraries were constructed by enzymatic fragmentation, end-repair, A-tailing, adaptor ligation, ligation cleanup, sample index PCR, and PCR cleanup.