To generate knockout mouse, ES cells were cultured and electroporated with the gene targeting vector. The genetically-modified cells were selected and transferred into embryos. The modified ES cells along with the cells of embryo were used to produce chimeric mice. Crosses between the chimeras and wild type mice were resulted heterozygous offspring.
Genticin
Genticin is a sterile and pyrogen-free solution of Gentamicin Sulfate, a broad-spectrum aminoglycoside antibiotic. It is intended for intramuscular, intravenous, or topical administration as a therapeutic agent.
Lab products found in correlation
2 protocols using genticin
Generation of Knockout Mice via ES Cells
To generate knockout mouse, ES cells were cultured and electroporated with the gene targeting vector. The genetically-modified cells were selected and transferred into embryos. The modified ES cells along with the cells of embryo were used to produce chimeric mice. Crosses between the chimeras and wild type mice were resulted heterozygous offspring.
Overexpressing MafA in ADMSCs
in 25 ml flasks 24 hours before the transfection. At day
10 of differentiation, cells were washed three times with
phosphate buffered saline (PBS, Calbiochem, Iran),
trypsinized, counted, and suspended at a density of 106/ml
in serum-free DMEM-HG. Then, 100 µl of cells were mixed
with 5 µg of suitable vector pCDNA(3.1+) in control group
and recombinant MafA/pCDNA 3.1(+) in the experimental
group in 0.4 ml electroporation cuvette (Biorad, USA), and
gently mixed by pipetting. The mixture was placed in an
electroporation system (GenePulser system II, Biorad, USA)
and one pulse of 140V was delivered for 15 milliseconds.
Following the electroporation, cells were plated onto a 25
ml flask that contained a differentiation medium and was
incubated at 37°C and 5% CO2. After 24 hours, Genticin
(350 µg/ml, Sigma, USA) was added to the growth media for
the positive selection of antibiotic-resistant ADMSCs. The
media were changed every 3 days, and Genticin selection
was maintained for 7 days. Antibiotic-resistant ADMSCs
were split to be grown for the differentiation.
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