Alexandrium tamarense microsatellite amplifications
of the three local strains with (A) specific primers ATB8 (lanes 1, 2 and 3) and
ATD8 (lanes 4, 5 and 6). Lanes 7 and 8 correspond to controls with primers
ATB8 without DNA. (B) Specific amplification with primers ATB1 (lanes 1, 2 and 3)
and ATF11 (lanes 6, 7 and 8). Lanes 4–5 and 9–10 are controls without
DNA for primer sets ATB1 and ATF11, respectively. M = 100-bp DNA size marker.
Species-specific microsatellite amplifications using A. catenella
and A. tamarense primers were carried out in a MaxiGene Gradiente
thermocycler (Axygen) in 1× PCR buffer, 20–50 ng of genomic DNA
template, 3 mM MgCl2, 100 µM each dNTP, 0.1 µM each primer
and 0.4 U of TopTaq DNA polymerase (Fermentas) in a 10-µL reaction volume.
Five microlitres of each PCR product were analysed in a 2 % agarose gel. A
Fermentas GeneRullerTM 100-bp DNA ladder was used for size estimation of
amplified fragments.