Fast isolation of microsatellite loci of very diverse repeat motifs by library enrichment in echinoderm species, Amphipholis squamata and Echinocardium cordatum

摘要

We conceived a microsatellite enrichment protocol in which probes of several repeat motifs are mixed during the whole procedure. The repeats found generally differed from those of the probes and were very diverse, from mononucleotides to pentanucleotides. We tested several modifications with alternative: (i) digestion/ligation buffers; (ii) polymerases; and (iii) purification methods. The simplest methods always worked as well as classical ones or even better, resulting in probably the simplest protocol for isolating microsatellites of diverse motifs from genomes with low microsatellite frequency. The proportion of positive clones, polymorphism levels and cross-specific amplification of microsatellite loci significantly vary between species.