GSM1973341: HCEC RNAseq, rep2; Homo sapiens; RNA-Seq
Cells of 1 to 3 million at 50-70% confluency were lysed by loading 1 ml of Tripure Isolation Reagent (Roche Diagnostics, 1667157). Two biological replica were prepared separately in each cell line. Total RNA was extracted by following manufacturer’s instruction. The extracted RNA was subjected on Bioanalyzer 2100 (Agilent Technologies) and RNA integrity number was confirmed as > 8. 4 ug each of total RNA was proceeded for sequence library preparation using Illumina TruSeq Stranded mRNA Library Prep Kit (Illumina, RS-122-2101). The quality and quantity of the library were assessed on Bioanalyzer 2100 and Qubit (Life Technologies). Total 4 libraries (2 x replica of HCEC, 2 x replica of HCT116) were pooled and sequenced on Illumina MiSeq using MiSeq Reagent Kit v2 (Illumina, MS-102-2002) generating 150 bp single-end read.