GSM8171932: T1-Tax-1; Homo sapiens; OTHER
Genomic DNA was isolated using Nucleospin Tissue kit (Macherey-Nagel, Germany) according to manufacturer's instructions. Next generation sequencing (NGS) libraries were prepared as described. Briefly, in the first PCR, 13.2 μg of gDNA was used as a template to achieve 250x coverage of the sgRNA library. In the second round, NGS (Illumina) adapters (stagger, index and index read primer sequences) were added at the ends of PCR amplicons. Products were run on 2% agarose gel, purified by using MN Nucleospin Gel and PCR Cleanup kit. NGS was performed at Genewiz (NJ, USA) by using Hiseq (Illumina) with at least 10 million reads/sample.