Inhibition testing
All DNA extracts were tested for inhibition by adding 3µL of extract
into a qPCR for the amplification of an internal positive control (DNA
derived from the plasmid pSD3). PCRs were set up in a total volume of 25
µL consisting of: 3 µL of extracted template DNA, 3 µL of internal
positive control DNA (0.08 ng/µl),1 µL of each primer/probe (0.2 µmol/L
forward primer; 0.4 µmol/L reverse primer; 0.1 µmol/L probe;
Supplemental file 2), 12.5 µL of TaqMan® Environmental Master Mix 2.0
(containing AmpliTaq GOLD DNA polymerase), and 3.5 µL ddH2O. PCR cycling
was as above.