A noninvasive molecular approach: exploiting species-locus-specific PCR primers in defeating numts and DNA cross-contamination of cercopithecidae

The lack of a standardized, noninvasive molecular approach to studying genetic aspects of primates has made it hard for primatologists to decode the evolutionary history of these species. Researchers must optimize their own techniques to fully exploit the available samples. Lack of species-locus-spe...

Full description

Bibliographic Details
Main Authors: Abu Bakar, Muhammad Abdul Latiff, Aifat, N. R., Yaakop, S., Md Zain, B. M.
Format: Article
Language:English
Published: Pakistan Agricultural Scientists Forum (PAS FORUM) 2017
Subjects:
Online Access:http://eprints.uthm.edu.my/4545/1/AJ%202017%20%28178%29%20A%20noninvasive%20molecular%20approach.pdf
_version_ 1825709864402288640
author Abu Bakar, Muhammad Abdul Latiff
Aifat, N. R.
Yaakop, S.
Md Zain, B. M.
author_facet Abu Bakar, Muhammad Abdul Latiff
Aifat, N. R.
Yaakop, S.
Md Zain, B. M.
author_sort Abu Bakar, Muhammad Abdul Latiff
collection UTHM
description The lack of a standardized, noninvasive molecular approach to studying genetic aspects of primates has made it hard for primatologists to decode the evolutionary history of these species. Researchers must optimize their own techniques to fully exploit the available samples. Lack of species-locus-specific primers also contributes to difficulties in using noninvasive genetic samples. Thus, the objectives of this study were to develop a standardized technique to collecting samples noninvasively, propose newly designed species-locus-specific primers, and optimize conditions for polymerase chain reaction (PCR) for Macaca fascicularis, M. nemestrina, Trachypithecus cristatus, and T. obscurus. Nine new species-locus-specific primers for three different loci of mitochondrial DNA, namely D-loop, cytochrome oxidase subunit I (COI), and cytochrome b, were successfully designed. These primers proved to be efficient in amplifying larger datasets (up to ~1,000 bp) of the targeted species in the optimized PCR conditions. The species-locus-specific primers are able to anneal to host DNA alone in highly contaminated feces of highlighted species. They can also offer alternatives measures in avoiding contamination related to nuclear insertion of mitochondrial pseudogenes (numts).
first_indexed 2024-03-05T21:48:52Z
format Article
id uthm.eprints-4545
institution Universiti Tun Hussein Onn Malaysia
language English
last_indexed 2024-03-05T21:48:52Z
publishDate 2017
publisher Pakistan Agricultural Scientists Forum (PAS FORUM)
record_format dspace
spelling uthm.eprints-45452021-12-07T06:50:00Z http://eprints.uthm.edu.my/4545/ A noninvasive molecular approach: exploiting species-locus-specific PCR primers in defeating numts and DNA cross-contamination of cercopithecidae Abu Bakar, Muhammad Abdul Latiff Aifat, N. R. Yaakop, S. Md Zain, B. M. TP Chemical technology The lack of a standardized, noninvasive molecular approach to studying genetic aspects of primates has made it hard for primatologists to decode the evolutionary history of these species. Researchers must optimize their own techniques to fully exploit the available samples. Lack of species-locus-specific primers also contributes to difficulties in using noninvasive genetic samples. Thus, the objectives of this study were to develop a standardized technique to collecting samples noninvasively, propose newly designed species-locus-specific primers, and optimize conditions for polymerase chain reaction (PCR) for Macaca fascicularis, M. nemestrina, Trachypithecus cristatus, and T. obscurus. Nine new species-locus-specific primers for three different loci of mitochondrial DNA, namely D-loop, cytochrome oxidase subunit I (COI), and cytochrome b, were successfully designed. These primers proved to be efficient in amplifying larger datasets (up to ~1,000 bp) of the targeted species in the optimized PCR conditions. The species-locus-specific primers are able to anneal to host DNA alone in highly contaminated feces of highlighted species. They can also offer alternatives measures in avoiding contamination related to nuclear insertion of mitochondrial pseudogenes (numts). Pakistan Agricultural Scientists Forum (PAS FORUM) 2017 Article PeerReviewed text en http://eprints.uthm.edu.my/4545/1/AJ%202017%20%28178%29%20A%20noninvasive%20molecular%20approach.pdf Abu Bakar, Muhammad Abdul Latiff and Aifat, N. R. and Yaakop, S. and Md Zain, B. M. (2017) A noninvasive molecular approach: exploiting species-locus-specific PCR primers in defeating numts and DNA cross-contamination of cercopithecidae. The Journal of Animal & Plant Sciences, 27 (3). pp. 1015-1053. ISSN 1018-7081
spellingShingle TP Chemical technology
Abu Bakar, Muhammad Abdul Latiff
Aifat, N. R.
Yaakop, S.
Md Zain, B. M.
A noninvasive molecular approach: exploiting species-locus-specific PCR primers in defeating numts and DNA cross-contamination of cercopithecidae
title A noninvasive molecular approach: exploiting species-locus-specific PCR primers in defeating numts and DNA cross-contamination of cercopithecidae
title_full A noninvasive molecular approach: exploiting species-locus-specific PCR primers in defeating numts and DNA cross-contamination of cercopithecidae
title_fullStr A noninvasive molecular approach: exploiting species-locus-specific PCR primers in defeating numts and DNA cross-contamination of cercopithecidae
title_full_unstemmed A noninvasive molecular approach: exploiting species-locus-specific PCR primers in defeating numts and DNA cross-contamination of cercopithecidae
title_short A noninvasive molecular approach: exploiting species-locus-specific PCR primers in defeating numts and DNA cross-contamination of cercopithecidae
title_sort noninvasive molecular approach exploiting species locus specific pcr primers in defeating numts and dna cross contamination of cercopithecidae
topic TP Chemical technology
url http://eprints.uthm.edu.my/4545/1/AJ%202017%20%28178%29%20A%20noninvasive%20molecular%20approach.pdf
work_keys_str_mv AT abubakarmuhammadabdullatiff anoninvasivemolecularapproachexploitingspecieslocusspecificpcrprimersindefeatingnumtsanddnacrosscontaminationofcercopithecidae
AT aifatnr anoninvasivemolecularapproachexploitingspecieslocusspecificpcrprimersindefeatingnumtsanddnacrosscontaminationofcercopithecidae
AT yaakops anoninvasivemolecularapproachexploitingspecieslocusspecificpcrprimersindefeatingnumtsanddnacrosscontaminationofcercopithecidae
AT mdzainbm anoninvasivemolecularapproachexploitingspecieslocusspecificpcrprimersindefeatingnumtsanddnacrosscontaminationofcercopithecidae
AT abubakarmuhammadabdullatiff noninvasivemolecularapproachexploitingspecieslocusspecificpcrprimersindefeatingnumtsanddnacrosscontaminationofcercopithecidae
AT aifatnr noninvasivemolecularapproachexploitingspecieslocusspecificpcrprimersindefeatingnumtsanddnacrosscontaminationofcercopithecidae
AT yaakops noninvasivemolecularapproachexploitingspecieslocusspecificpcrprimersindefeatingnumtsanddnacrosscontaminationofcercopithecidae
AT mdzainbm noninvasivemolecularapproachexploitingspecieslocusspecificpcrprimersindefeatingnumtsanddnacrosscontaminationofcercopithecidae