GSM9290004: MDA ETV4 KO (replicate 3); Homo sapiens; RNA-Seq
For harvesting of the 3D cultures, collagen 1 gels were washed once with HBSS and digested with 1ml collagenase I at 300U/ml for 10 minutes at 37°C. After 10 minutes, mechanical disruption of the gels was performed by pipetting. Additional 5 minutes incubation at 37°C was performed. The suspension was then washed with HBSS by centrifugation and cell pellet was frozen at -80°C. RNA extraction was performed using RNeasy Mini Kit (Qiagen); DNA was removed by DNase treatment. RNA-seq libraries for deep-sequencing were prepared with TruSeq Stranded mRNA (Illumina)