title: GSM9054806: NC1, non-transplanted hearts of negative control, scRNAseq; Mus musculus; RNA-Seq
library_construction_protocol: To generate single-cell suspensions, fresh heart tissues were cut into 1 mm3 pieces, and were digested by 0.5 mg/mL liberase solution  at 37 °C for 20 minutes. To ensure full digestion of tissues, heart tissue pieces were gently shaken in a 37 °C shaker during the process. After digestion, the supernatant medium was harvested and add the same volume of Dulbecco's Modified Eagle's medium (DMEM) containing 10% fetal bovine serum (FBS) to terminate the digestive reaction, new liberase solution was added in residual tissue pieces for further digestion. The mixture of supernatant and DMEM was then filtered through a 40 μm cell strainer to obtain single cell suspension. The collected cells were centrifuged at 300 g/min at 4°C for 5 minutes. Subsequently, cell pellets were suspended in PBS or DEME for further experiments. Single cell library was generated using Chromium Single Cell 3' Reagent Kit v2 and v3 chemistry (10X Genomics) following standard protocol. Library was generated and sequenced on a NovaSeq platform (Illumina) with paired-end 150 bp (PE 150) sequencing strategy.