Abstract
This article documents the addition of 83 microsatellite marker loci and 96 pairs of single-nucleotide polymorphism (SNP) sequencing primers to the Molecular Ecology Resources Database. Loci were developed for the following species: Bembidion lampros, Inimicus japonicus, Lymnaea stagnalis, Panopea abbreviata, Pentadesma butyracea, Sycoscapter hirticola and Thanatephorus cucumeris (anamorph: Rhizoctonia solani). These loci were cross-tested on the following species: Pentadesma grandifolia and Pentadesma reyndersii. This article also documents the addition of 96 sequencing primer pairs and 88 allele-specific primers or probes for Plutella xylostella.
Highlights
Linkers were ligated to the digested DNA, and the resulting fragments were enriched for microsatellites by hybridization to and magnetic capture of biotinylated repeat probes
Enriched genomic fragments captured by streptavidin-coated magnetic beads were amplified by PCR, ligated to Roche/454 Titanium Multiplex Identifier (MID) adapters and size fractionated in an agarose gel
We primarily focused on finding tetranucleotide repeats as they typically provide clear allele assignment
Summary
Abbreviata specimen from San Matías gulf (Patagonia, Argentina), using the BioSprint DNA Blood Kit (Qiagen, Hilden, Germany), according to manufacturer’s tissue protocol. Five micrograms of extracted genomic DNA were completely digested with a restriction enzyme (five-base cutter) that generated blunt-end fragments.
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