title: HiSeq X Ten paired end sequencing; RNA seq in U-937 cells wildtype clones and rs2431697 region deletion clones
design: RNA seq in U-937 cells wildtype clones and rs2431697 region deletion clones
library_construction_protocol: we deleted a 30-bp region at rs2431697 in the U-937 cell line using CRISPR/Cas9 with two sgRNAs targeted around the rs2431697 site. Plasmids expressing Cas9 and sgRNAs were nucleofected into cells, and cell clones were screened to obtain bi-allelic 30-bp deletion clones. Clones exposed to CRISPR/Cas9 complexes with wildtype genotype were selected as controls.  Total RNA was extracted using TRIzol reagent (15596018, Thermo Fisher)  1 μg total RNA was purified to remove rRNA and was used to generate the cDNA libraries using Illumina NEBNext® Ultra™ Directional RNA Library Prep Kit