title: Illumina HiSeq 4000 paired end sequencing; RNA Seq of control and IRGM KD HT-29 Cells
design: RNA Seq of control and IRGM KD HT-29 Cells
library_construction_protocol: HT-29 control and IRGM KD cells were collected from 60mm dish RNA was extracted from HT29 cells(three biological replicates) using RNeasy mini kit (QIAGEN, #74104). The quality and quantity of total RNA was checked using agarose gel and Qubit 3.0 After assessing the quality of RNA, 800-900 ng of total RNA was subjected to NEBNext® ultra™ directional RNA library prep kit for Illumina® (#E7420) using NEBNextPoly(A) mRNA magnetic isolation module (NEB #E7490). The QC of the prepared library was performed using High-Sensitivity Tape Station Kit (Agilent 2200, 5067-5585 (reagents) #5067-5584 ( screen tapes)) for fragment length distribution and Qubit dsDNA HS assay kit (Invitrogen, Q32851) for quantifying the library