Multiplex pre‐amplification for noninvasive genetic sampling: is the extra effort worth it?
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 perfor...
Published in: | Molecular Ecology Resources |
---|---|
Main Authors: | , |
Format: | Article in Journal/Newspaper |
Language: | English |
Published: |
Wiley
2010
|
Subjects: | |
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 |
id |
crwiley:10.1111/j.1755-0998.2009.02818.x |
---|---|
record_format |
openpolar |
spelling |
crwiley:10.1111/j.1755-0998.2009.02818.x 2023-12-03T10:31:32+01:00 Multiplex pre‐amplification for noninvasive genetic sampling: is the extra effort worth it? DE BARBA, M. WAITS, L. P. 2010 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 en eng Wiley http://onlinelibrary.wiley.com/termsAndConditions#vor Molecular Ecology Resources volume 10, issue 4, page 659-665 ISSN 1755-098X 1755-0998 Genetics Ecology, Evolution, Behavior and Systematics Biotechnology journal-article 2010 crwiley https://doi.org/10.1111/j.1755-0998.2009.02818.x 2023-11-09T14:35:01Z 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. Article in Journal/Newspaper Ursus arctos Wiley Online Library (via Crossref) Molecular Ecology Resources 10 4 659 665 |
institution |
Open Polar |
collection |
Wiley Online Library (via Crossref) |
op_collection_id |
crwiley |
language |
English |
topic |
Genetics Ecology, Evolution, Behavior and Systematics Biotechnology |
spellingShingle |
Genetics Ecology, Evolution, Behavior and Systematics Biotechnology DE BARBA, M. WAITS, L. P. Multiplex pre‐amplification for noninvasive genetic sampling: is the extra effort worth it? |
topic_facet |
Genetics Ecology, Evolution, Behavior and Systematics Biotechnology |
description |
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. |
format |
Article in Journal/Newspaper |
author |
DE BARBA, M. WAITS, L. P. |
author_facet |
DE BARBA, M. WAITS, L. P. |
author_sort |
DE BARBA, M. |
title |
Multiplex pre‐amplification for noninvasive genetic sampling: is the extra effort worth it? |
title_short |
Multiplex pre‐amplification for noninvasive genetic sampling: is the extra effort worth it? |
title_full |
Multiplex pre‐amplification for noninvasive genetic sampling: is the extra effort worth it? |
title_fullStr |
Multiplex pre‐amplification for noninvasive genetic sampling: is the extra effort worth it? |
title_full_unstemmed |
Multiplex pre‐amplification for noninvasive genetic sampling: is the extra effort worth it? |
title_sort |
multiplex pre‐amplification for noninvasive genetic sampling: is the extra effort worth it? |
publisher |
Wiley |
publishDate |
2010 |
url |
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 |
genre |
Ursus arctos |
genre_facet |
Ursus arctos |
op_source |
Molecular Ecology Resources volume 10, issue 4, page 659-665 ISSN 1755-098X 1755-0998 |
op_rights |
http://onlinelibrary.wiley.com/termsAndConditions#vor |
op_doi |
https://doi.org/10.1111/j.1755-0998.2009.02818.x |
container_title |
Molecular Ecology Resources |
container_volume |
10 |
container_issue |
4 |
container_start_page |
659 |
op_container_end_page |
665 |
_version_ |
1784257845165817856 |