characteristics: ERC000011
characteristics: ENA-CHECKLIST
characteristics: 2022-10-26T00:21:09Z
characteristics: ENA-FIRST-PUBLIC
characteristics: 2022-10-26T00:21:09Z
characteristics: ENA-LAST-UPDATE
characteristics: SAMEA10774142
characteristics: External Id
characteristics: Duke-NUS Medical School
characteristics: INSDC center name
characteristics: 2022-10-26T00:21:09Z
characteristics: INSDC first public
characteristics: 2022-10-26T00:21:09Z
characteristics: INSDC last update
characteristics: public
characteristics: INSDC status
characteristics: ERS8425404
characteristics: SRA accession
characteristics: E-MTAB-11119:Dengue virus mutant
characteristics: Submitter Id
characteristics: ArrayExpress
characteristics: broker name
characteristics: Huh7.5.1
characteristics: cell line
characteristics: hepatocyte
characteristics: cell type
characteristics: human
characteristics: common name
characteristics: Protocols: A pool of CRISPR mutagenized cells resisting a dengue virus mutant cytolytic infection was harvested from a live-dead genetic screen. 2 million resistant cells were harvested from each sub-library, and subjected to total genomic DNA extraction using DNA mini kit (Qiagen) according to the manufacturerвЂ™s instructions The sgRNA were amplified from gDNA in two-step PCR using Herculase II Fusion DNA Polymerase (Agilent). For first PCR, 20 ug of gDNA for each library was amplified for 16 cycles. For 2nd PCR, 1 reaction containing 5ul PCR product for each sub-library was amplified for 27 cycles, using Illumina indexed primers. PCR products were gel purified (Qiagen gel purification kit) and sequenced on Illumina MiSeq platform.
characteristics: description
characteristics: adult
characteristics: developmental stage
characteristics: hepatocellular carcinoma
characteristics: disease
characteristics: genome-wide CRISPR KO library under phenotypic selection
characteristics: genetic modification
characteristics: wild type genotype
characteristics: genotype
characteristics: Dengue virus mutant
characteristics: infect
characteristics: Homo sapiens
characteristics: organism
characteristics: Homo sapiens
characteristics: organism
characteristics: liver
characteristics: organism part
characteristics: Homo sapiens
characteristics: scientific_name
characteristics: male
characteristics: sex
characteristics: Dengue virus mutant
characteristics: title