6. Extraction and Sequencing of sgRNA or Barcodes from Genomic DNA

Identification of sgRNA, shRNA, or barcode sequences in experimental cell or tissue samples from screens with pooled libraries requires (1) isolation and amplification of the target sgRNA or barcode inserts from the genomic DNA, and (2) Next-Generation Sequencing (NGS) of amplified sgRNA or barcode sequences using an Illumina NGS instrument. Following amplification, representation levels of each of the sgRNA or barcode sequences are then quantified to assess enrichment or depletion relative to the starting library or relevant control samples, depending on the type of screen.