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Simple Protocol for Molecular Fingerprinting of Human Oral Microbiota Samples in Lab Classes

    Authors: Ana C. Henriques1, Paolo De Marco1,*
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    Affiliations: 1: CESPU, Instituto de Investigação e Formação Avançada em Ciências e Tecnologias da Saúde (IINFACTS), 4585-116 Gandra PRD, Portugal
    AUTHOR AND ARTICLE INFORMATION AUTHOR AND ARTICLE INFORMATION
    • Received 02 February 2017 Accepted 27 October 2017 Published 26 January 2018
    • ©2018 Author(s). Published by the American Society for Microbiology
    • [open-access] This is an Open Access article distributed under the terms of the Creative Commons Attribution-Noncommercial-NoDerivatives 4.0 International license (https://creativecommons.org/licenses/by-nc-nd/4.0/ and https://creativecommons.org/licenses/by-nc-nd/4.0/legalcode), which grants the public the nonexclusive right to copy, distribute, or display the published work.

    • *Corresponding author. Mailing address: CESPU, Instituto de Investigação e Formação Avançada em Ciências e Tecnologias da Saúde (IINFACTS), Rua Central de Gandra, 1317, 4585-116, Gandra PRD, Portugal
    Source: J. Microbiol. Biol. Educ. January 2018 vol. 19 no. 1 doi:10.1128/jmbe.v19i1.1305
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    Abstract:

    DNA fingerprinting is a major tool in identifying individuals and in evidence matching. However, this technique can be difficult to reproduce in practical classes. Here, we report on distinct PCR profiles obtained when amplifying saliva DNA of a score of distinct individuals with Random Amplified Polymorphic DNA (RAPD)-PCR primer BOXA1R. The RAPD-PCR method is simple and efficient for discrimination between bacterial strains and is used in this instance to obtain personalized fingerprints of each individual’s oral microbiota. We present real results with undergraduate students confirming that this procedure is easily feasible in practical classes. Based on the results presented, we suggest a laboratory activity for undergraduate Molecular Biology or Microbiology students.

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2018-01-26
2018-07-16

Abstract:

DNA fingerprinting is a major tool in identifying individuals and in evidence matching. However, this technique can be difficult to reproduce in practical classes. Here, we report on distinct PCR profiles obtained when amplifying saliva DNA of a score of distinct individuals with Random Amplified Polymorphic DNA (RAPD)-PCR primer BOXA1R. The RAPD-PCR method is simple and efficient for discrimination between bacterial strains and is used in this instance to obtain personalized fingerprints of each individual’s oral microbiota. We present real results with undergraduate students confirming that this procedure is easily feasible in practical classes. Based on the results presented, we suggest a laboratory activity for undergraduate Molecular Biology or Microbiology students.

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Figures

Image of FIGURE 1

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FIGURE 1

Results obtained by five groups of students (A to E). A) 1, 2, 3, 4 = individual saliva DNA samples. 5 = water negative control. B) 1, 3, 4, 5, 6 = individual saliva DNA samples. 7 = water negative control. 2 = same sample as nº 1, loaded twice by mistake. C) 1, 2, 3, 4, 5 = individual saliva DNA samples. 6 = water negative control. D) 1, 2, 3, 4, 5 = individual saliva DNA samples. 6 = water negative control. E) 1, 2, 3, 4 = individual saliva DNA samples. 5 = water negative control. M = molecular weight marker.

Source: J. Microbiol. Biol. Educ. January 2018 vol. 19 no. 1 doi:10.1128/jmbe.v19i1.1305
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Image of FIGURE 2

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FIGURE 2

Histogram of grades (0 to 20) from 67 students. 0 to 7 (red) = not proficient; 8 to 15 (blue) = proficient; 16 to 20 (green) = highly proficient. Questions and students’ answer examples can be found in Appendix 6 , with our suggestion for a scoring rubric linked to student learning objectives. Authorization for using students’ grades was obtained from our internal Ethics Committee.

Source: J. Microbiol. Biol. Educ. January 2018 vol. 19 no. 1 doi:10.1128/jmbe.v19i1.1305
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