title: GSM2460416: Control sgRNA rep3; Homo sapiens; RNA-Seq
library_construction_protocol: Prior to sample preparation for RNA-seq, dead cells were removed on day 3 after puromycin selection (day 5 after lentiviral transduction of sgRNAs) using Dead Cell Removal Kit (Miltenyi Biotec #130-090-101, San Diego, CA) following the manufacturer’s manual. Total RNAs were subsequently isolated using PureLink RNA Mini Kit (Thermo Fisher Scientific) following the manufacturer’s manual. An in-column DNA digestion step was included to remove any residual genomic DNA contamination. To construct RNA-seq libraries, 500 ng total RNA was used for polyA mRNA-selection using NEBNext Poly(A) mRNA Magnetic Isolation Module (#E7490, New England Biolabs, Inc.  Ipswich, MA), followed by library construction via NEBNext Ultra RNA Library Prep Kit for Illumina (#E7530, New England Biolabs, Inc., Ipswich, MA).
attributes.tag: GEO Accession
attributes.value: GSM2460416