Multiplex pre‐amplification for noninvasive genetic sampling: is the extra effort worth it?

• Published in: Molecular Ecology Resources
• Main Authors: DE BARBA, M., WAITS, L. P.
• Format: Article in Journal/Newspaper
• Language: English
• Published: Wiley 2010
• Subjects: Genetics; Ecology, Evolution, Behavior and Systematics; Biotechnology; Ursus arctos
• Online Access: http://dx.doi.org/10.1111/j.1755-0998.2009.02818.x; https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1111%2Fj.1755-0998.2009.02818.x; https://onlinelibrary.wiley.com/doi/pdf/10.1111/j.1755-0998.2009.02818.x; https://onlinelibrary.wiley.com/doi/full-xml/10.1111/j.1755-0998.2009.02818.x

Abstract Microsatellite genotyping of hair and faeces using standard polymerase chain reaction (PCR) resulted in low success rates and high error rates in a 2003–2004 pilot study using noninvasive genetic sampling for the brown bear ( Ursus arctos ) in the Italian Alps. Thus, we evaluated the performance of multiplex pre‐amplification for improving microsatellite genotyping results. Brown bear faecal DNA extracts of varying quality ( n = 33) and hair DNA extracts of poor ( n = 32) and good ( n = 34) quality were used to compare standard PCR and pre‐amplification. In contrast to previous studies, there was no significant difference between methods for individual locus amplification success, genotyping error and genotyping success rates for scat and hair samples. The use of pre‐amplification requires an additional investment of time and resources, and our results raise questions about the universal value of pre‐amplification approaches. We suggest that researchers carefully evaluate the performance of pre‐amplification compared to standard PCR using field‐collected samples from the study area of interest before engaging in large‐scale noninvasive genetic analyses.