WT_T0_B
Homo sapiens
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.
disease
lung adenocarcinoma
organism
Homo sapiens
organism
Homo sapiens
collection date
not collected
scientific_name
Homo sapiens
common name
human
organism part
lung
developmental stage
adult
genotype
wild type genotype
isolate
not applicable
cell_line
HCC827
cell_type
epithelial cell
geographic location (country and/or sea)
not collected
ENA-LAST-UPDATE
2023-12-14