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Bacterial DNA Extraction Using Individual Enzymes and Phenol/Chloroform Separation

    Authors: Mitchell Henry Wright1,*, Joseph Adelskov2, Anthony Carlson Greene2
    VIEW AFFILIATIONS HIDE AFFILIATIONS
    Affiliations: 1: Division of Environmental and Biomolecular Systems, Institute of Environmental Health, Oregon Health & Science University, Portland, OR 97239-3098; 2: School of Natural Sciences, Griffith University, Nathan Campus, Queensland, Australia
    AUTHOR AND ARTICLE INFORMATION AUTHOR AND ARTICLE INFORMATION
    • Received 02 May 2017 Accepted 03 July 2017 Published 01 September 2017
    • ©2017 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.

    • Supplemental materials available at http://asmscience.org/jmbe
    • *Corresponding author. Mailing address: Division of Environmental and Biomolecular Systems, Institute of Environmental Health, Oregon Health & Science University, 3181 S.W. Sam Jackson Park Rd., Portland, OR, 97239-3098. Phone: 503-346-3434. E-mail: wrigmit@ohsu.edu.
    Source: J. Microbiol. Biol. Educ. September 2017 vol. 18 no. 2 doi:10.1128/jmbe.v18i2.1348
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    Abstract:

    Marmur (4) developed one of the first detailed comprehensive methods for purifying bacterial DNA. This procedure is now outdated, and can be difficult to follow for those with limited experience in molecular biology. Here, we provide a modernized, simplified protocol for extracting bacterial DNA and discuss how this can be incorporated into microbiology laboratory courses for biology majors.

Key Concept Ranking

Bacterial Cell Wall
0.6623455
Sodium Dodecyl Sulfate
0.5248334
Agarose Gel Electrophoresis
0.48006326
16s rRNA Sequencing
0.41046953
0.6623455

References & Citations

1. Wright MH, Sirdaarta J, White A, Greene AC, Cock IE2017GC-MS headspace analysis of Terminalia ferdinandiana fruit and leaf extracts which inhibit Bacillus anthracis growthPharmacogn J91738210.5530/pj.2017.1.14 http://dx.doi.org/10.5530/pj.2017.1.14
2. Wright MH, Farooqui SM, White AR, Greene AC2016Production of manganese oxide nanoparticles by Shewanella speciesAppl Environ Microbiol82175402540910.1128/AEM.00663-16273425594988204 http://dx.doi.org/10.1128/AEM.00663-16
3. Zhang Y, Li S, Gan R, Zhou T, Xu D, Li H2015Impacts of gut bacteria on human health and diseasesInt J Mol Sci1647493751910.3390/ijms16047493258496574425030 http://dx.doi.org/10.3390/ijms16047493
4. Marmur J1961A procedure for the isolation of deoxyribonucleic acid from micro-organismsJ Mol Bio3220821810.1016/S0022-2836(61)80047-8 http://dx.doi.org/10.1016/S0022-2836(61)80047-8
5. Adelskov J, Patel BKC2016A molecular phylogenetic framework for Bacillus subtilis using genome sequences and its application to Bacillus subtilis subspecies stecoris strain D7XPN1, an isolate from a commercial food-waste degrading bioreactor3 Biotech69610.1007/s13205-016-0408-8283301664809863 http://dx.doi.org/10.1007/s13205-016-0408-8
6. Amaro A, Duarte E, Amado A, Ferronha H, Botelho A2008Comparison of three DNA extraction methods for Mycobacterium bovis, Mycobacterium tuberculosis and Mycobacterium avium subspAvium Letter Appl Micr47181110.1111/j.1472-765X.2008.02372.x http://dx.doi.org/10.1111/j.1472-765X.2008.02372.x
7. Ogg CD, Patel BKC2009Caloramator australicus sp. nov., a thermophilic, anaerobic bacterium from the Great Artesian Basin of AustraliaInt J Sys Evol Micr599510110.1099/ijs.0.000802-0 http://dx.doi.org/10.1099/ijs.0.000802-0
8. Emmert EAB2013Biosafety guidelines for handling microorganisms in the teaching laboratory: development and rationaleJ Microbiol Biol Educ141788310.1128/jmbe.v14i1.531238583563706168 http://dx.doi.org/10.1128/jmbe.v14i1.531
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/content/journal/jmbe/10.1128/jmbe.v18i2.1348
2017-09-01
2017-11-20

Abstract:

Marmur (4) developed one of the first detailed comprehensive methods for purifying bacterial DNA. This procedure is now outdated, and can be difficult to follow for those with limited experience in molecular biology. Here, we provide a modernized, simplified protocol for extracting bacterial DNA and discuss how this can be incorporated into microbiology laboratory courses for biology majors.

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FIGURE 1

Agarose gel containing λHind III linear standard (A) and purified genomic DNA (B, C). Visualized under UV light.

Source: J. Microbiol. Biol. Educ. September 2017 vol. 18 no. 2 doi:10.1128/jmbe.v18i2.1348
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