Journal: PLoS ONE
Article Title: Comparative Analysis of the Recently Discovered hAT Transposon TcBuster in Human Cells
Figure Lengend Snippet: The effect of transposon and transposase plasmid dose on the number of drug-resistant colonies formed. ( a ) HEK-293 cells in 6-well plates were transfected in triplicate with either 12.5 ng (light grey bars), 50 ng (dark grey bars), or 500 ng (black bars) of pTcBNeo carrying the neomycin-resistance transposon and 0 ng, 100 ng, 250 ng, or 500 ng of pCMV- TcBuster expressing the transposase. ( b ) HEK-293 cells in 6-well plates were transfected in triplicate with 500 ng of pTcBNeo plasmid carrying the neomycin-resistance transposon and the indicated amount of pCMV- TcBuster (0.5 ng, 1 ng, 5 ng, 10 ng, 25 ng, 50 ng, 100 ng, 250 ng, or 500 ng). In both a and b , pUC19 was used as filler DNA to increase the total amount of DNA transfected to 1 µg. Cells were diluted 1∶750 in selection media and grown for two weeks to allow drug-resistant cells to multiply and form colonies. The colonies were fixed, stained, and counted. The mean and standard error of the mean (SEM; n = 3) are shown.
Article Snippet: Plasmid constructs All plasmids were prepared for assays by endotoxin-free maxiprep (Qiagen, Valencia, CA). pUC19 (Invitrogen, Carlsbad, CA) was used as filler plasmid DNA for the FuGene6 transfections.
Techniques: Plasmid Preparation, Transfection, Expressing, Selection, Staining