GSM8194104: B16-BL6 cells, sgNDUFS4, rep2; Mus musculus; RNA-Seq
RNA samples were prepared with TRIzol® Reagent. Briefly, cells in 60 mm dish with ~70% confluent cells were incubated with 1 ml TRIzol® Reagent for 5 minutes and then transferred into a 1.5 ml tube and mixed with 0.2 ml chloroform. After vigorously shaking for 15 seconds, samples were centrifuged at 12000 g for 15 minutes at 4 °C. The aqueous phage was collected and mixed with an equal volume of 100% isopropanol. The mixture was then centrifuged at 12000 g for 15 minutes at 4 °C to obtain RNA pellet. The supernatant was removed, and the obtained RNA pellet was washed with 75% ethanol 3 times. RNA pellet was dry and resuspended into RNase free water. The concentration of RNA was measured with a nanodrop spectrophotometer, and the quality of RNA was determined by agarose gel electrophoresis. Samples were processed by Genewiz in accordance with their internal protocols for paired end strand independent sequencing