title: GSM8686096: WT_HCT116_1; Homo sapiens; RNA-Seq
library_name: GSM8686096
library_construction_protocol: Total RNA was extracted using TRIzol reagent (Invitrogen), followed by further purification using PureLink RNA Mini Kit (Cat No. 12183018A;Invitrogen) with DNase treatment (QIAGEN). RNA quality and quantity were analyzed with 2100 Expert (Agilent). RIN (the RNA Integrity Number) of all samples were above 8.0. This protocol involves the removal of ribosomal RNA (rRNA) using biotinylated, target-specific oligos combined with Ribo-Zero rRNA removal beads. 12 RNA-seq samples were pooled and sequenced on HiSeq using Illumina TruSeq Stranded Total RNA Library Preparation kit and paired-end sequencing. The RNA is fragmented into small pieces and the cleaved RNA fragments are copied into first strand cDNA using reverse transcriptase and random primers, followed by second strand cDNA synthesis using DNA Polymerase I and RNase H. The resulting double-strand cDNA is used as the input to a standard Illumina library prep with end-repair, adapter ligation and PCR amplification being performed to give you a sequencing ready library. The final purified product is then quantitated by qPCR before cluster generation and paired end sequencing on HiSeq 4000.