title: GSM4873811: LC_UVR_24hr_1_RNA-seq; Mus musculus; RNA-Seq
library_construction_protocol: Ears were split and floated upon Trypsin-GNK (2.5mg/ml trypsin (Sigma), 7.35mg/ml NaCl, 0.35mg/ml KCl, 0.85mg/ml glucose) for 1 hr at 37 degrees C. Epidermis was separated and digested in trypsin-GNK with 100 ug/ml DNase I (Roche) and 1X Dnase buffer (1.21 mg/ml Tris base, 0.5mg/ml MgCl2, 73ug/ml CaCl2) for 20 min at 37 degrees C with agitation. Isolated cells were washed in medium and stained with Live/Dead Fixable Near-IR Dye (Molecular Probes), followed by mouse Fc block (BD) and anti-EpCAM-PE, anti-Sca1-FITC, anti-CD3-PerCPCy5.5, anti-CD45-APC, anti-TCRd-BV421, anti-MHC-II(I-A/I-E)-BV605. Cells were then washed and sorted on a BD FACS Aria Fusion directly into buffer RLT+. RNA was isolated from sorted cells using RNeasy Micro Plus Kit (Qiagen). cDNA was prepared with Ovation RNA-Seq System V2 (NuGEN) and libraries were constructed with Ultralow Library System V2 (NuGEN). Resulting libraries were pooled and sequenced on an Illumina HiSeq 2500 platform with single ended 75 bp reads.
attributes.tag: GEO Accession
attributes.value: GSM4873811