GSM8782820: Nanos-Upd tumor Replicate 2; Drosophila melanogaster; RNA-Seq
Tesis tumors were dissected in sterile Schnieder's media supplemented with 10% fetal bovine serum. Testis tissue was digested to completion in TrypLE supplemented with 2mg/mL collagenase by incubating with intermittent agitation at 37C. Digested tissue was filtered first through at 40um filter, then a 10um filter before pelleting cells by centrifugation at 1,200rpm. After removing the supernatant cells were washed once in HBSS followed by centrifugation. After the second wash, cells were resuspended in HBSS and cell count/viability was obtained with Trypan blue. Library preparation for RNA-seq was performed following the instructions for 10X Genomics Chromium Next GEM Single Cell 3'Kit v3.1.