GSM3131831: A549.starr_seq.dex.1hr.rep5; Homo sapiens; OTHER
Each washed cell pellet harvested from a 500 cm2 plate was lysed in a 50 mL conical tube with 5 mL of a 1:10 mixture of β-mercaptoethanol and Qiagen Buffer RLT. The lysates were vortexed, snap frozen, and stored at -80°C. Lysates were thawed on ice and passed through a 20-guage needle 10x. Total RNAs were purified with the Qiagen RNeasy midi-columns performing the on-column DNase step. The amount of purified RNA was quantified using a Nanodrop instrument, and RNA quality was assessed using an Agilent TapeStation 2200 with RNA ScreenTape. After adding 1 ul RNase Block (Agilent) samples were stored at -80 Total RNAs harvested from A549 cells transfected with the GM12878 Whole Genome STARR-seq plasmid library were subjected to poly-A enrichment, DNase digestion, reporter-specific reverse transcription, and PCR enrichment.