Figure 2

Episomal detection using PCR. A. PCR reaction products from 8 individual transgenic adult D. melanogaster (Canton S) with the autonomous element Hermes 7011 (lanes 1-8). Lanes 9-12 contain the products of identical PCR reactions using the equal amounts of genomic DNA from non-transgenic Canton S individuals. The diagram below schematically illustrates the structure and size of PCR products arising from Hermes elements with their terminal inverted repeats (thick arrows) joined as shown using the primers indicated (66R, 2681F) with variable (n) numbers of nucleotides of intercalary DNA. The positions and size in basepairs of molecular weight standards are indicated. B. PCR reaction products from 9 individual M. domestica adults from 3 geographically distinct natural populations. The diagram below schematically illustrates the structure and size of PCR products arising from Hermes elements with their terminal inverted repeats (thick arrows) joined as shown using the primers indicated (66R, 2431F) with variable (n) numbers of nucleotides of intercalary DNA. Molecular weight markers (m) and their sizes in basepairs are shown. Roman numerals refer to bands that were excised, reamplified, cloned and sequenced. The results are shown in Table 5. C. PCR reaction products from 6 individual M. domestica adults from the laboratory colony, Cs. DNA from unfertilized eggs (e) from this colony was also used as template in these PCR reactions. The positions and size in basepairs of molecular weight standards are indicated.