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Bi-allelic gene targeting in mouse embryonic stem cells.

Methods (San Diego, Calif.) (2011-02-04)
Peri H Tate, William C Skarnes
PMID21288739
ABSTRACT

The EUCOMM and KOMP programs have generated targeted conditional alleles in mouse embryonic stem cells for nearly 10,000 genes. The availability of these stem cell resources will greatly accelerate the functional analysis of genes in mice and in cultured cells. We present a method for conditional ablation of genes in ES cells using vectors and targeted clones from the EUCOMM and KOMP conditional resources. Inducible homozygous cells described here provide a precisely controlled experimental system to study gene function in a model cell.

MATERIALS
Product Number
Brand
Product Description
S9763
Sigma-Aldrich
Sodium phosphate dibasic, ACS reagent, ≥99.0%
S9638
Sigma-Aldrich
Sodium phosphate monobasic monohydrate, ACS reagent, ≥98%
G7776
Sigma-Aldrich
Glutaraldehyde solution, Grade I, 70% in H2O, specially purified for use as an electron microscopy fixative or other sophisticated use
D4551
Sigma-Aldrich
N,N-Dimethylformamide, for molecular biology, ≥99%
G7528
Sigma-Aldrich
D-(+)-Glucose, ≥99.5% (GC), BioXtra
I8896
Sigma-Aldrich
IGEPAL® CA-630, for molecular biology
E6511
Sigma-Aldrich
Ethylenediaminetetraacetic acid tetrasodium salt dihydrate, BioReagent, suitable for cell culture, 98.5-102.0%