24-7-GFP-POS0
Protocols: After a minimum of 72 hrs post nucleofection DNA or RNA was extracted (DNA Quick extract, Epicenter or Qiagen Allprep DNA/RNA mini, #80204) or cells were harvested for further analysis by respectively PCR, qPCR or flow cytometry analysis/FACS sorting. K562 cells were cultured in RPMI 1640 + L-glutamine (Hyclone SH30027.LS), 10% fetal calf serum (FCS, Gibco) and 50 U/mL penicillin and 50 μg/mL streptomycin (Gibco 15140-163). Cells were incubated at 37°C and 5% CO2 and passaged every 3-4 days to 100.000 cells/ml. CRISPR-Cas9 technology was used to make mutations or deletions in the regions described in the results section. After a minimum of 72 hrs post nucleofection DNA or RNA was extracted (DNA Quick extract, Epicenter or Qiagen Allprep DNA/RNA mini, #80204) or cells were harvested for further analysis by respectively PCR, qPCR or flow cytometry analysis/FACS sorting. The first PCR was performed using Q5 polymerase (NEB), the second nested PCR with KAPA HiFi HotStart Ready mix (Roche).
ENA first public
2021-07-26
ENA last update
2021-07-26
External Id
SAMEA7741262
INSDC center alias
Department of Hematology, Erasmus University Medical Center
INSDC center name
Department of Hematology, Erasmus University Medical Center
INSDC first public
2021-07-26T12:17:16Z
INSDC last update
2021-07-26T12:17:16Z
INSDC status
public
Submitter Id
E-MTAB-9937:24-7-GFP-POS0
broker name
ArrayExpress
cell line
K562
cell type
myeloblast
common name
human
developmental stage
adult
disease
adult acute myeloid leukemia
genotype
t(3;8)
organism part
bone marrow cell
phenotype
GFP positive sorted
sample name
E-MTAB-9937:24-7-GFP-POS0
treatment
CRISPR EVI1 PR CTCF sgRNA#1