GSM5574491: RNASEH1_scramble_rep2; Homo sapiens; RNA-Seq
Total RNAs from cultured cells were extracted with Trizol Reagent (Life Technologies) according to the manufacturer's protocol. RNAs were further treated with DNase I (Ambion, DNA-free kit) and cDNAs were reverse transcribed with PrimeScript II RTase (TAKARA) at 42 ℃ for 2hr, and then applied for qPCR analysis using the SYBR Green Realtime PCR Master Mix (TOYOBO). For RNA-seq samples from PA1 cells treated with scramble or RNase H1 shRNAs, ribo minus RNA libraries were prepared using Illumina TruSeq Stranded Total RNA LT Sample Prep Kit according to the manufacturer's protocol with slight modifications in the step for synthesizing first strand cDNA, which used PrimeScript enzyme mix (TAKARA) for reverse transcription at 42 ℃ for 2 hr. All libraries were size-selected with AmpureXP beads (Agencourt) and quantified using Agilent Bio analyzer 2100 and Qubit high-sensitivity dsDNA assay.