alias: GSM4543995
title: GSM4543995: Cas9-positive initial (adherent) condition replicate 3; Homo sapiens; RNA-Seq
library_construction_protocol: At each time point indicated (either L0, T0, or Tf), cells were harvested and genomic DNA was extracted using QIAmp Blood Maxi Kits (QIAGEN #51194). Genomic encoded sgRNA sequences were PCR amplified with two rounds of PCR to increase abundance of the initial sgRNA population and insert sample barcodes necessary for identification of condition and replicate number. PCR products were gel purified, quantitated by Qubit (Invitrogen) and pooled prior to high throughput sequencing. Pooled preparation containing all barcoded samples were sequenced using an Illumina NextSeq 550 75-cycle high output kit (#20024906). FASTQ files were obtained containing raw reads for all samples and were demultiplexed to obtain individual FASTQ files for each sample: one initial pooled sgRNA library file (L0), 3x Cas9-positive initial condition replicates (T0), 3x Cas9-positive final condition replicates (Tf), 3x Cas9-negative initial condition replicates (T0), 3x Cas9-negative final condition replicates (Tf).
identifiers.namespace: GEO
identifiers.id: GSM4543995