title: MCF10A PT T1
organism: Homo sapiens
description: Protocols: Inducible SpCas9 expressing cells were seeded for 1000x representation per sgRNA, and infected at a multiplicity of infection (MOI) of 0.3, to avoid multiple sgRNA infections per cell, using the previously published and validated genome-wide human lentiviral library (Kosuke Yusa Human GW CRISPR guide RNA library V1). After puromycin selection (1 µg/mL), cells were exposed to doxycycline for 3 days and harvested at day 0 (T0) and on day 14 (T1). Genomic DNA was extracted from the T=0 and T=1 samples using DNeasy Blood and Tissue Kit (Qiagen) as per manufacturers instructions. sgRNA sequence were amplified with Q5 polymerase (NEB) with the following primers Forward 5'- ACACTCTTTCCCTACACGACGCTCTTCCGATCTCTTGTGGAAAGGACGAAACA and Reverse 5'- TCGGCATTCCTGCTGAACCGCTCTTCCGATCTCTAAAGCGCATGCTCCAGAC.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2023-05-22
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2023-05-22
attributes.tag: External Id
attributes.value: SAMEA113428052
attributes.tag: INSDC center alias
attributes.value: The institute of Cancer Research
attributes.tag: INSDC center name
attributes.value: The institute of Cancer Research
attributes.tag: INSDC first public
attributes.value: 2023-05-22T00:14:50Z
attributes.tag: INSDC last update
attributes.value: 2023-05-22T00:14:50Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-12939:MCF10A PT T1
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell_line
attributes.value: MCF-10A
attributes.tag: cell_type
attributes.value: epithelial cell
attributes.tag: common name
attributes.value: human
attributes.tag: genotype
attributes.value: TP53-/-
attributes.tag: isolate
attributes.value: not applicable
attributes.tag: organism part
attributes.value: breast
attributes.tag: sample name
attributes.value: E-MTAB-12939:MCF10A PT T1
attributes.tag: scientific_name
attributes.value: Homo sapiens