RNA-seq of human K562 cell line, CRISPR/cas9 clone K562:SCR5
Single-cell derived clones from CRISPR/cas9 knockout, Scramble gRNA treated sample--no editing, wildtype control. Total RNA was isolated with the Qiagen miRNeasy Mini Kit. NEBNext Ultra RNA Library Prep Kitfor Illumina by following manufacturers recommendations (NEB, Ipswich, MA, USA). Briefly,mRNA were first enriched with Oligod(T) beads. Enriched mRNAs were fragmented for 15minutes at 94 C. First strand and second strand cDNA were subsequently synthesized. cDNAfragments were end repaired and adenylated at 3ends,and universal adapter was ligated tocDNA fragments, followed by index addition and library enrichment with limited cycle PCR.