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A PCR Method That Can Be Further Developed into PCR-RFLP Assay for Eight Animal Species Identification
There are many PCR-based methods for animal species identification; however, their detection numbers are limited or could not identify unknown species. We set out to solve this problem by developing a universal primer PCR assay for simultaneous identification of eight animal species, including goat,...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Hindawi
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5832126/ https://www.ncbi.nlm.nih.gov/pubmed/29629212 http://dx.doi.org/10.1155/2018/5890140 |
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author | Guan, Feng Jin, Yu-Ting Zhao, Jin Xu, Ai-Chun Luo, Yuan-Yuan |
author_facet | Guan, Feng Jin, Yu-Ting Zhao, Jin Xu, Ai-Chun Luo, Yuan-Yuan |
author_sort | Guan, Feng |
collection | PubMed |
description | There are many PCR-based methods for animal species identification; however, their detection numbers are limited or could not identify unknown species. We set out to solve this problem by developing a universal primer PCR assay for simultaneous identification of eight animal species, including goat, sheep, deer, buffalo, cattle, yak, pig, and camel. In this assay, the variable lengths of mitochondrial DNA were amplified using a pair of universal primers. PCR amplifications yielded 760 bp, 737 bp, 537 bp, 486 bp, 481 bp, 464 bp, 429 bp, and 359 bp length fragments for goat, sheep, deer, buffalo, cattle, yak, pig, and camel, respectively. This primer pair had no cross-reaction with other common domestic animals and fish. The limit of detection varied from 0.01 to 0.05 ng of genomic DNA for eight animal species in a 20 µl PCR mixture. Each PCR product could be further digested into fragments with variable sizes and qualitative analysis by SspI restriction enzyme. This developed PCR-RFLP assay was sufficient to distinguish all targeted species. Compared with the previous published related methods, this approach is simple, with high throughput, fast processing rates, and more cost-effective for routine identification of meat in foodstuffs. |
format | Online Article Text |
id | pubmed-5832126 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Hindawi |
record_format | MEDLINE/PubMed |
spelling | pubmed-58321262018-04-08 A PCR Method That Can Be Further Developed into PCR-RFLP Assay for Eight Animal Species Identification Guan, Feng Jin, Yu-Ting Zhao, Jin Xu, Ai-Chun Luo, Yuan-Yuan J Anal Methods Chem Research Article There are many PCR-based methods for animal species identification; however, their detection numbers are limited or could not identify unknown species. We set out to solve this problem by developing a universal primer PCR assay for simultaneous identification of eight animal species, including goat, sheep, deer, buffalo, cattle, yak, pig, and camel. In this assay, the variable lengths of mitochondrial DNA were amplified using a pair of universal primers. PCR amplifications yielded 760 bp, 737 bp, 537 bp, 486 bp, 481 bp, 464 bp, 429 bp, and 359 bp length fragments for goat, sheep, deer, buffalo, cattle, yak, pig, and camel, respectively. This primer pair had no cross-reaction with other common domestic animals and fish. The limit of detection varied from 0.01 to 0.05 ng of genomic DNA for eight animal species in a 20 µl PCR mixture. Each PCR product could be further digested into fragments with variable sizes and qualitative analysis by SspI restriction enzyme. This developed PCR-RFLP assay was sufficient to distinguish all targeted species. Compared with the previous published related methods, this approach is simple, with high throughput, fast processing rates, and more cost-effective for routine identification of meat in foodstuffs. Hindawi 2018-02-05 /pmc/articles/PMC5832126/ /pubmed/29629212 http://dx.doi.org/10.1155/2018/5890140 Text en Copyright © 2018 Feng Guan et al. https://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Article Guan, Feng Jin, Yu-Ting Zhao, Jin Xu, Ai-Chun Luo, Yuan-Yuan A PCR Method That Can Be Further Developed into PCR-RFLP Assay for Eight Animal Species Identification |
title | A PCR Method That Can Be Further Developed into PCR-RFLP Assay for Eight Animal Species Identification |
title_full | A PCR Method That Can Be Further Developed into PCR-RFLP Assay for Eight Animal Species Identification |
title_fullStr | A PCR Method That Can Be Further Developed into PCR-RFLP Assay for Eight Animal Species Identification |
title_full_unstemmed | A PCR Method That Can Be Further Developed into PCR-RFLP Assay for Eight Animal Species Identification |
title_short | A PCR Method That Can Be Further Developed into PCR-RFLP Assay for Eight Animal Species Identification |
title_sort | pcr method that can be further developed into pcr-rflp assay for eight animal species identification |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5832126/ https://www.ncbi.nlm.nih.gov/pubmed/29629212 http://dx.doi.org/10.1155/2018/5890140 |
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