Quick Extract (Lucigen QE09050): Protocol according to manufacturers instructions. [lentiCRISPR bulk]  gDNA was extracted and sonicated to ~1000bp. sgRNA containing fragments were recovered with biotinylated RNA probes targeting the flanking region on the lenticrispr v2 backbone and pulled down with streptavidin beads. gDNA fragments were PCRed first with primers flanking the sgRNA and second with Illumina sequencing adapters. [targeted microRaft]  gDNA was extracted with in QuickExtract buffer and input into a PCR reaction containing barcoded primers that flank the gRNA insert. This PCR was cleaned up over a DNA purification column, and uniquely-barcoded products were pooled into a second PCR reaction using illumina sequencing primers.
GEO Accession
GSM4146796