Conditions for PCR amplification were: 2 min at 95 uC,
followed by 35 cycles, each consisting of 95 uC for 40s, 50–52 uC for
40s, and 1–3 min (according to the examined region size) at 72 uC.
The final PCR extension step was 10 min at 72 uC. PCR products
were analysed on a 1% agarose gel and visualized with specific dyes
for DNA. Purified PCR products were sequenced (using BigDye
terminator v3.1; Applied Biosystems) on both strands using the
same PCR primers. Cycle-sequencing reactions were sequenced on an
Applied Biosystems ABI Prism 3100 DNA Sequencer