title: KO_T15_B
organism: Homo sapiens
description: Protocols: 80 million HCC827 cells (WT or RBM10-KO) were collected at each timepoint, snap-frozen in liquid nitrogen, and kept in -80°C prior to genomic DNA extraction. Genomic DNA extraction was performed with Blood & Cell Culture DNA Maxi Kit (Qiagen) according to the manufacturer's instructions. gRNA inserts were amplified using two round of PCR reactions. First, a 600bp region of the lentiviral library vectors containing the sgRNA region was amplified using ordinary PCR with Q5 high-fidelity DNA polymerase for 25 cycles. A total of 120μg gDNA was used as template for each sample (60reactions: 2μg gDNA/50μl reaction). Second, the amplified PCR product was subjected to another PCR reaction with Illumina next-generation sequencing adaptors for 10 cycles and 200bp product was amplified.
attributes.tag: disease
attributes.value: lung 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: lung
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: genotype
attributes.value: RBM10_KO
attributes.tag: isolate
attributes.value: not applicable
attributes.tag: cell_line
attributes.value: HCC827
attributes.tag: cell_type
attributes.value: epithelial cell
attributes.tag: geographic location (country and/or sea)
attributes.value: not collected
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2023-12-14