title: cell_culture_1
organism: Homo sapiens
description: Protocols: 6.3x10e6 HEK293_M2 cells harboring the virally integrated human ORFeome version 3.1 collection were thawed and cultured for 1 day to leave the cells time to recover. Genomic DNA was recovered from 16x10e6 cells and 6 micrograms of gDNA were used to amplify the collection (3 PCR reactions with 2 micrograms of gDNA). Cells were lysed using SNET buffer (10mM Tris pH 8.0, 0.1M EDTA, 0.5% SDS, 0.1mg/ml Proteinase K, 25microgram/ml RNAse A). Genomic DNA was isolated using 1 volume of Phenol/Chloroform/Isoamyl Alcohol (25:24:1) pH 8.0, precipitated by adding 2 volumes of 95% Ethanol (-20C), washed with ice cold 70% Ethanol and resuspended in Tris-HCl pH 8.0. Following PCR, the amplified ORFs were used directly to prepare a Nextera sequencing library (Illumina Nextara DNA Sample Preparation Kit)
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2014-05-15T17:02:21Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2018-03-08T17:22:00Z
attributes.tag: External Id
attributes.value: SAMEA2464865
attributes.tag: INSDC center name
attributes.value: University of Toronto
attributes.tag: INSDC first public
attributes.value: 2014-05-15T17:02:21Z
attributes.tag: INSDC last update
attributes.value: 2018-03-08T17:22:00Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-2498:cell_culture_1
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell line
attributes.value: HEK293
attributes.tag: common name
attributes.value: human
attributes.tag: genetic modification
attributes.value: Transduced cells (lentivirus)
attributes.tag: genotype
attributes.value: HEK293 with human ORFeome version 3.1
attributes.tag: sample name
attributes.value: E-MTAB-2498:cell_culture_1
attributes.tag: scientific_name
attributes.value: Homo sapiens
attributes.tag: specimen with known storage state
attributes.value: fresh specimen