FIG. 7. Strategy for high-throughput plasmid transfer detection.

Plasmid-containing donor (D) and recipient (R) cells are mixed (it can be done at different ratios), filtered in a 96-filter-well plate (0.22 µm) and placed on a layer of solid conjugation medium (containing 5 µg/ml DNase I). After 4 h at 37°C, selection for transconjugants (T) is applied by adding antibiotic-containing medium and application of gentle vacuum. Thus, for each well the transfer frequencies can be assessed by a number of methods, such as plating on selective medium, fluorescent confocal microscopy (gfp-expressing plasmids), LacZ measurements, or quantitative PCR (Modified from (123)).