8_128322614n
Protocols: RNP complexes were formed incubating sgRNA and Cas9 (IDT) for 20-30 at RT before nucleofection using the Neon (Thermofischer) with buffer R using program 3. The MUTZ3 cell lines were cultured in αMEM (HyClone) with 20% fetal calf serum (FCS) and 20% conditioned 5637 medium. The 293T were cultured in DMEM (Gibco) with 10% FCS. All cell lines were supplemented with 50 U/mL penicillin and 50 μg/mL streptomycin. Viable frozen AML cells were thawed and resuspended in IMDM medium supplemented with: 20% BIT medium (Stem cell technologies), 1x β-mercaptoethanol (1000x Life technologies), 6 µg/ml LDL (Sigma Aldrich), human IL6, IL3, G-CSF, GM-CSF at 20 ng/ml and FLT3, SCF at 50 ng/ml (Peprotech. Genomic DNA was extracted 48 or 72 h after transfection using Quick Extract buffer (Epicenter) For amplicon sequencing we used the Illumina PCR-based custom amplicon sequencing method using the TruSeq Custom Amplicon index kit (Illumina). The first PCR was performed using Q5 polymerase (NEB), the second nested PCR with KAPA HiFi HotStart Ready mix (KapaBiosystems).
ENA first public
2021-07-26
ENA last update
2021-07-26
External Id
SAMEA7742529
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-9945:8_128322614n
broker name
ArrayExpress
cell line
MUTZ-3
cell type
myeloblast
common name
human
developmental stage
adult
disease
acute myeloid leukemia
genotype
inv(3)
organism part
bone marrow
sample name
E-MTAB-9945:8_128322614n