Simple sequence repeat anchored polymerase chain reaction
amplification (SSR-PCR) is a genetic typing technique based on primers anchored
at the 5' or 3' ends of microsatellites, at high primer annealing
temperatures. This technique has already been used in studies of genetic
variability of several organisms, using different primer designs. In order
to conduct a detailed study of the SSR-PCR genomic targets, we cloned and
sequenced 20 unique amplification products of two commonly used primers,
CAA(CT)
6
and (CA)
8
RY, using
Biomphalaria glabrata
genomic DNA as template. The sequences obtained were novel
B. glabrata
genomic sequences. It was observed that 15 clones contained microsatellites
between priming sites. Out of 40 clones, seven contained complex sequence
repetitions. One of the repeats that appeared in six of the amplified fragments
generated a single band in Southern analysis, indicating that the sequence
was not widespread in the genome. Most of the annealing sites for the CAA(CT)
6
primer contained only the six repeats found within the primer sequence.
In conclusion, SSR-PCR is a useful genotyping technique. However, the premise
of the SSR-PCR technique, verified with the CAA(CT)
6
primer, could not be supported since the amplification products did not
result necessarily from microsatellite loci amplification.
