An optimisation approach to increase DNA amplification success of otter faeces

摘要: Faeces have proved to be a suitable non-invasive DNA source for microsatellite analysis in wildlife research. For the success of such studies it is essential obtain highest possible PCR amplification rate. These rates are still relatively low most carnivorous species, especially otter (Lutra lutra). We therefore optimised entire genotyping process by combining our findings with results from previous gain high rate reliable genotypes. investigated influence faecal quality relation quantity slimy secretions and three levels storage periods at −20°C on success. Further, we tested cost-effective time-saving Chelex extraction method against profitable QIAamp® Stool Kit (Qiagen), compared methods - standard single-step protocol, single-locus two-step procedure multiplex regarding errors. The (median: 94%; mean: 78%) was achieved using samples consisting only jelly extracted Mini (Qiagen) immediately after collection amplified following time cost efficient protocol. procedure, also referred as pre-amplification approach, turned out main improvement increases about 11% reduces errors 53%, notably allelic dropouts.