GSM7520280: gRNA sequencing -  Late Pluripotent Biol Rep2; Mus musculus; OTHER
Cells were dissociated and sorted based on BFP, SSEA1-APC and X-GFP expression using a BD FACSAria II SORP. Genomic DNA was extracted from cell pellets using the DNeasy Blood and Tissue Kit (Qiagen, 69504). For amplification of the gRNAs and introduction of the Illumina-sequencing adapters, two consecutive PCRs were performed by using the Q5 High-Fidelity DNA Polymerase (NEB, M0491), adding the Illumina adapsters for NGS. Single-end sequencing (1 x 50 bp) on an Illumina HiSeq 2500. Each sample contains two technical replicates