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Fast and reliable DNA extraction protocol for identification of species in raw and processed meat products sold on the commercial market

Pavel Espinoza Alvarado
  • Departamento de Ciencias Agronómicas y Veterinarias, Instituto Tecnológico de Sonora, Av. Antonio Caso S/N Colonia Villa ITSON, Ciudad Obregón, Sonora, México
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Ramón Miguel Molina Barrios
  • Departamento de Ciencias Agronómicas y Veterinarias, Instituto Tecnológico de Sonora, Av. Antonio Caso S/N Colonia Villa ITSON, Ciudad Obregón, Sonora, México
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Javier Arturo Munguía Xóchihua
  • Departamento de Ciencias Agronómicas y Veterinarias, Instituto Tecnológico de Sonora, Av. Antonio Caso S/N Colonia Villa ITSON, Ciudad Obregón, Sonora, México
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Juan Francisco Chávez Hernández
  • Corresponding author
  • Departamento de Ciencias Agronómicas y Veterinarias, Instituto Tecnológico de Sonora, Av. Antonio Caso S/N Colonia Villa ITSON, Ciudad Obregón, Sonora, México
  • Email
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
Published Online: 2017-09-02 | DOI: https://doi.org/10.1515/opag-2017-0051

Abstract

In this work a protocol for the extraction of DNA from the meat of different animals (beef, pork, and horse) was established. The protocol utilized TE lysis buffer with varying concentrations of phenol and chloroform as a base reagent. Reactions were carried out for verying time periods and under differing temperatures. All samples analyzed were obtained from commercial grade meat sourced from the local region. 12 samples were used for methodological optimization with 30 repetitions per sample. Once optimized, purity results for the three species were 1.7 with a concentration (determined spectrophotometrically at 260 nm) of 100 μl/ml of DNA. The protocol was tested using 465 different meat samples from different animal species. All meat used was fresh and processed. Results showed a purity of 1.35 ± 0.076 and a DNA concentration of 70 ± 0.31 μl for a time duration of 1.5 hours. These results were tested by polymerase chain reaction (PCR) as reported by several authors. The extracts were tested using different PCR reactions using specific primers for horses. Results suggest that there was 39 positive samples. The proposed methodology provides an efficient way to detect DNA concentration and purity, suitable for amplification with PCR.

Keywords : DNA extraction Standardization meat products

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About the article

Received: 2016-12-02

Accepted: 2017-05-03

Published Online: 2017-09-02

Published in Print: 2017-08-28


Citation Information: Open Agriculture, Volume 2, Issue 1, Pages 469–472, ISSN (Online) 2391-9531, DOI: https://doi.org/10.1515/opag-2017-0051.

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© 2017. This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License. BY-NC-ND 4.0

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