title: GSM3764380: AI4_MDA-MB-231_Anillin knockout sgRNA4; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA was extracted from either control, or anillin deficient (two different CRISPR probes, referred hereafter as CRISPR1 and CRISPR2) MDA-MB-231 cell lines using mirVana miRNA Isolation Kit (ThermoFisher Scientific, Cat #AM1560). The RNA quality was determined by Bioanalyzer (Agilent, Santa Clara, CA). To isolate the polyA RNA, NEBNext Poly(A) mRNA Magnetic Isolation Module (New England BioLabs, Ipswich, MA) combined with SMARTer Apollo NGS library preparation system (Takara, Mountain View, CA) were used with a total of 1 µg of good quality total RNA as input. The NEBNext Ultra Directional RNA Library Prep Kit (New England BioLabs) was then used for polyA RNA library preparation, which is dUTP-based stranded library. The library was indexed and amplified under PCR cycle number of 11. After library Bioanalyzer QC analysis and quantification, individually indexed and compatible libraries were proportionally pooled and sequenced using Illumina HiSeq 1000 sequencer.