title: GSM8513932: Healthy control 2; Homo sapiens; RNA-Seq
library_construction_protocol: Patients included were adults and had a diagnosis of histologically proven lGVHD from the dermatology and hematology departments of Saint Louis Hospital, Paris, France. Skin biopsies were obtained using 4 mm punch and normal skin were obtained from healthy donors undergoing breast plastic surgery. Skin biopsies were transferred in NaCl 9‰ and immediately digested for 4h30 at 37°C using whole skin dissociation kit (Miltenyi) according to the manufacturer protocol. Single cell suspension obtained was then processed with Asteria kit (Scipio Bioscience) with 30,000 cells input in 2 kits for each sample to generate 3' enriched cDNA with corresponding cell barcode. The cell barcoded cDNA was fragmented, end-repaired then adapter was ligated to the extremities using NEBNext Ultra II FS DNA libary kit for illumina. Selective amplification of cDNA fragment containing the cell barcode was done with custum P5-R1 library primer from Asteria kit and indexed P7-R2 primier from NEBNext Mulitplex Oligos kit. Libraries were sequenced on the illumina NovaSeq 6000 sequencer to generate paired-end read. 25 bases for read1 and 70 bases for read2. read1 contains cell barcode and read2 contains 3' sequence of cDNA.