title: GSM7245829: sample2_Melanoma; Homo sapiens; RNA-Seq
library_construction_protocol: After surgical resection, tumor specimens were washed with antibiotic-containing media and minced with crossed scalpels under sterile conditions. Tissue was dissociated via enzymatic digestion using 1,500 U/ml collagenase IV (Gibco/Life Technologies), 1,000 U/ml hyaluronidase (Sigma), and 0.05 mU/ml DNase IV (Gibco) in RPMI for 1 h at 37°C followed by mechanical agitation. Centrifugation over Ficoll-Hypaque gradient was used to separate debris from the single cell suspension. Finally, cells were cryopreserved in 10% DMSO + bovine serum. RNA was then purified from cells using RNeasy Micro Kits (Qiagen). RNA libraries were prepared for sequencing using the SMART-Seq v4 for CDNA and the Nextera XT kit for library creation on a NextSeq 550 instrument