GSM5237189: RNA-seq L8 rep1; Homo sapiens; RNA-Seq
For RNA-seq samples, cells were lysed in RLT buffer supplemented with 550 μM 2-Mercaptoethanol, gDNA was removed, and RNA was purified on spin columns using the RNeasy Plus kit (Qiagen, #74134) according to kit directions. Libraries were prepared from 1μg RNA with the NEBNext Ultra RNA Library Prep Kit for Illumina (NEB; #E7530L). Briefly, mRNA was purified from total RNA using poly-T oligo magnetic beads, fragmented, and reverse transcribed to double stranded cDNA. Fragments were end-repaired, blunt-ended, 3'adenylated, and ligated to hairpin loop NEBNext Adaptors. 150~200 bp cDNA fragments were selected, PCR-amplified, and sequenced on an Illumina Novaseq 6000 platform, generating 150bp paired-end reads. For RNA-seq samples, libraries were prepared from 1μg RNA with the NEBNext Ultra RNA Library Prep Kit for Illumina (NEB; #E7530L). Briefly, mRNA was purified from total RNA using poly-T oligo magnetic beads, fragmented, and reverse transcribed to double stranded cDNA. Fragments were end-repaired, blunt-ended, 3'adenylated, and ligated to hairpin loop NEBNext Adaptors. 150~200 bp cDNA fragments were selected, PCR-amplified, and sequenced on an Illumina Novaseq 6000 platform, generating 150bp paired-end reads.