G418 is an aminoglycoside antibiotic , the molecular genetic tests , is the most commonly used stably transfected resistance screening reagents. By inhibiting the transposon Tn601, Tn5 gene ribosome interference function block protein synthesis, such as prokaryotic and eukaryotic cells to produce toxins , including bacteria , yeast, plant and mammalian cells , including protozoa and worms. When the neo gene was integrated into the DNA of eukaryotic cells , the neo gene is able to start the coding sequence is transcribed into mRNA, which acquired resistance to aminoglycoside phosphotransferase product efficient expression , the cells which acquired resistance selection in G418 containing of the growth medium . This selection features G418 , has been in gene transfer , gene knockout , transgenic animals resistance screening and other aspects can be widely applied .
Shipped at room temperature , dry storage at room temperature , soluble in water , methanol , and the solution stored at -20 ℃ eukaryotic cell lines because each different sensitivity to G418 , or the lines of the new cell lines stably transfected with non- stable transfection Kill cells required amount will need to experiment optimized. Optimum dosage curve obtained through the establishment of cell death . The cells were diluted to 1000cell/ml, adding 100ul per well of 24-well plates with the medium , the G418 concentration in each well diluted to 0,100,200,300,400,500,600,700,800,900,1000,1100 ug / ml in 12 levels , cultured for 10-14 days in all cells death lowest concentration as a benchmark, usually around 400-800 , screening than this concentration no matter how high a level to maintain the use of filtering half the concentration can be.
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From:Lactic acid bacteria
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