title: GSM4626277: RNA-Seq-Clone21-CTCF&TSS-CRISPR#1-rep2; Homo sapiens; RNA-Seq
library_construction_protocol: HAP1 cells were washed with PBS, and resuspended in 1mL TRIzol (Thermo Fisher), with 200µL chloroform then added. RNA was extracted using RNeasy Mini Kit (Qiagen). For sample ID #s 1732-1737, sequencing libraries were constructed from 500 ng of DNase-treated, total RNA using the ScriptSeqv2 Complete Kit (Illumina BHMR1224). Later samples (ID #s 2564-2565, 2694-2697), Illumina Truseq Stranded Total RNA kit was used, as ScriptSeqv2 was discontinued. Briefly, the RNA was depleted of rRNA using the Ribo-Zero removal reagents and fragmented. First strand cDNA was then synthesized using a 5' tagged random hexamer and reverse transcription, followed by annealing of a 5' tagged, 3'-end blocked terminal-tagged oligo and second strand synthesis. The Di-tagged cDNA fragments were purified, barcoded, and PCR-amplified for 15 cycles. Libraries were quality checked and quantified prior to 2 x 76 bp sequencing on the Illumina NextSeq 500. rRNA-depleted RNA
attributes.tag: GEO Accession
attributes.value: GSM4626277