title: GSM5957519: Sham_control; Mus musculus; RNA-Seq
library_construction_protocol: The electroporated area was punched out with a circular 4 mm biopsy punch, and subsequently minced with fine scissors to small pieces (approximately 1 mm2). The same, but non-injured area was used in control livers. The resulting fragments were further processed by enzymatic digestion in 5 mL enzyme mix consisting of dispase (50 caseinolytic units/ml), collagenase (2 mg/ml), and DNase (30 μg/ml), for 30 min at 37 °C under constant agitation (180 RPM). Enzyme activity was inhibited by adding 5 ml of phosphate-buffered saline (PBS) supplemented with 10% fetal bovine serum (FBS). Dissociated cells in suspension were passed through a 70 μm strainer and centrifuged at 500 × g for 5 min at 4 °C. Red blood cell lysis (Thermo Fisher 00-4333-57) was performed for 2 min and stopped with 10% FBS in PBS. 1 ng of pre-amplified cDNA from an estimated 1000 cells was tagmented by Nextera XT (Illumina) with a custom P5 primer (Integrated DNA Technologies). Single cell libraries were sequenced in a 100 bp paired-end run on the Illumina HiSeq4000 using 0.2 nM denatured sample and 5% PhiX spike-in.