organism: Homo sapiens
description: Protocols: >1000-fold coverage of mutagenized cells (Brunello library) was infected with PeV-A1 or PeV-A2 at MOI 0.1. Resistant cell colonies were harvested. Uninfected starting populations of mutagenized cells were used as the unselected reference. Total genomic DNA from both virus-resistant and uninfected cells was respectively extracted using QIAamp DNA Mini Kit (Qiagen). A first round of PCR was used to amplify the guide RNA sequences encoded in the gDNA, followed by a second round of PCR to add the barcodes/adapters for amplicon sequencing. 2% agarose gels and a QIAquick gel extraction kit were used to purify the amplicons.
attributes.tag: disease
attributes.value: Intestinal Type Adenocarcinoma
attributes.tag: organism
attributes.value: Homo sapiens
attributes.tag: organism
attributes.value: Homo sapiens
attributes.tag: collection date
attributes.value: not collected
attributes.tag: scientific_name
attributes.value: Homo sapiens
attributes.tag: common name
attributes.value: human
attributes.tag: organism part
attributes.value: colon
attributes.tag: genotype
attributes.value: lentiCas9-Blast; lentiGuide-Puro; SLC35A1/EXTL3 knockout, PeV-A1 infection
attributes.tag: isolate
attributes.value: not applicable
attributes.tag: cell_line
attributes.value: HT-29
attributes.tag: cell_type
attributes.value: epithelial cell
attributes.tag: geographic location (country and/or sea)
attributes.value: not collected
attributes.tag: infect
attributes.value: PeV-A1