GSM5918073: PA-1 DGCR8 heterozygote chromatin_associated replicate2; Homo sapiens; RNA-Seq
The chromatin-associated RNA was subsequently DNAse treated using RQ1 DNAse (Promega) according to manufacturer's instructions at a total volume of 100 µl. After 15 minutes of DNAse incubation at 37 °C, the solution was mixed with equal volume of Acid-Phenol:Chloroform (ThermoFisher) and vortexed until the solution turned white. After centrifuging at maximum speed in a cooled centrifuge, 100 µl of the top aqueous solution was mixed with 250 µl Ethanol and 10 µl 3M sodium acetate pH 5.2 (ThermoFisher). The mixture was then spun down at maximum speed in a cooled centrifuge prior to removing the supernatant. The RNA pellet was then washed with 70% Ethanol, spin centrifuged for 5 minutes in a cool centrifuge at maximum speed. The RNA pellet was then air dried and resuspended in H2O. After extraction, ≥ 200 ng of total RNA was used for rRNA removal stranded library sequencing with DNBseq PE100 by BGI Genomics.