title: GSM4873641: VP64_rep1_HbF_high; Homo sapiens; OTHER
library_construction_protocol: genomic DNA was extracted using the Qiagen Blood and Tissue kit. sgRNA integrant PCR to amplify spacer sequences was performed with Herculase II reaction buffer (1X), forward and reverse primers (0.5 μM each), dimethyl sulfoxide (DMSO) (8%), deoxynucleotide triphosphates (dNTPs) (0.25 μM each), Herculase II Fusion DNA Polymerase (0.5 reactions) using the following cycling conditions: 95°C for 2 min; 20 cycles of 95°C for 15 s, 60°C for 20 s, 72°C for 30 s; 72°C for 5 min. Multiple reactions of no more than 200 ng each were used to amplify from 6.6 ug gDNA (~1x106 cell genomes) per pool. Samples were subjected to additional rounds of PCR to add sequencing adaptors and indices before Illumina sequencing. sgRNA integrant PCR to amplify spacer sequences was performed with Herculase II reaction buffer (1X), forward and reverse primers (0.5 μM each), dimethyl sulfoxide (DMSO) (8%), deoxynucleotide triphosphates (dNTPs) (0.25 μM each), Herculase II Fusion DNA Polymerase (0.5 reactions) using the following cycling conditions: 95°C for 2 min; 20 cycles of 95°C for 15 s, 60°C for 20 s, 72°C for 30 s; 72°C for 5 min. Multiple reactions of no more than 200 ng each were used to amplify from 6.6 ug gDNA (~1x106 cell genomes) per pool. Samples were subjected to additional rounds of PCR to add sequencing adaptors and indices before Illumina sequencing.
attributes.tag: GEO Accession
attributes.value: GSM4873641