GSM8962904: CTFL-GFPplus; Mus musculus; RNA-Seq
Dorsal skin (around 2g) was placed in HBSS in 10cm dishes and transferred in digestion medium (1 U/mL Dispase II, 0,8 U/ml Liberase DH, 10U/mL DNase in Hanks' Balanced Salt Solution solution) during 15 min at 37C. The dorsal skin was scraped and cell solution was incubated under agitation during 15 min at 37C. This two-step was repeated once and total cell solution was inhibited with DMEM with 10% FBS. Cells were washed, LEC were isolated as CD31 and GFP positive cells by FACS analysis Single-cells were captured using the Chromium Controller and the Chromium Next GEM single cell 3' v3.1 kit (10x Genomics, Inc.). The library was profiled with the HS NGS kit for the Fragment Analyzer (Agilent Technologies) and quantified using the KAPA library quantification kit (Roche Diagnostics). The library was sequenced on the Illumina NextSeq550 instrument using the High Output 150 cycles kit (Illumina). The sequencing parameters were 28, 10, 10, 90 cycles (read 1, index 1, index 2, read 2). A minimum of 125406 mean reads/cell were obtained