Bacterial DNA Extraction Using Individual Enzymes and Phenol/Chloroform Separation
†

Mitchell Henry Wright; Joseph Adelskov; Anthony Carlson Greene

doi:10.1128/jmbe.v18i2.1348

Bacterial DNA Extraction Using Individual Enzymes and Phenol/Chloroform Separation †

Authors:

Mitchell Henry Wright^1,*, Joseph Adelskov², Anthony Carlson Greene²

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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: [email protected].

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 This procedure is now outdated, and can be difficult to follow for those with limited experience in molecular biology.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 IE 2017 GC-MS headspace analysis of Terminalia ferdinandiana fruit and leaf extracts which inhibit Bacillus anthracis growth Pharmacogn J 9 1 73 82 10.5530/pj.2017.1.14 http://dx.doi.org/10.5530/pj.2017.1.14

2. Wright MH, Farooqui SM, White AR, Greene AC 2016 Production of manganese oxide nanoparticles by Shewanella species Appl Environ Microbiol 82 17 5402 5409 10.1128/AEM.00663-16 27342559 4988204 http://dx.doi.org/10.1128/AEM.00663-16

3. Zhang Y, Li S, Gan R, Zhou T, Xu D, Li H 2015 Impacts of gut bacteria on human health and diseases Int J Mol Sci 16 4 7493 7519 10.3390/ijms16047493 25849657 4425030 http://dx.doi.org/10.3390/ijms16047493

4. Marmur J 1961 A procedure for the isolation of deoxyribonucleic acid from micro-organisms J Mol Bio 3 2 208 218 10.1016/S0022-2836(61)80047-8 http://dx.doi.org/10.1016/S0022-2836(61)80047-8

5. Adelskov J, Patel BKC 2016 A 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 bioreactor 3 Biotech 6 96 10.1007/s13205-016-0408-8 28330166 4809863 http://dx.doi.org/10.1007/s13205-016-0408-8

6. Amaro A, Duarte E, Amado A, Ferronha H, Botelho A 2008 Comparison of three DNA extraction methods for Mycobacterium bovis, Mycobacterium tuberculosis and Mycobacterium avium subsp Avium Letter Appl Micr 47 1 8 11 10.1111/j.1472-765X.2008.02372.x http://dx.doi.org/10.1111/j.1472-765X.2008.02372.x

7. Ogg CD, Patel BKC 2009 Caloramator australicus sp. nov., a thermophilic, anaerobic bacterium from the Great Artesian Basin of Australia Int J Sys Evol Micr 59 95 101 10.1099/ijs.0.000802-0 http://dx.doi.org/10.1099/ijs.0.000802-0

8. Emmert EAB 2013 Biosafety guidelines for handling microorganisms in the teaching laboratory: development and rationale J Microbiol Biol Educ 14 1 78 83 10.1128/jmbe.v14i1.531 23858356 3706168 http://dx.doi.org/10.1128/jmbe.v14i1.531

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Supplemental Material

Appendix 1: Pre-laboratory questions, Appendix 2: Pre-laboratory answer key, Appendix 3: DNA extraction flowchart, Appendix 4: Agarose gel preparation for electrophoresis
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2017-09-01

2019-10-22

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Author and Article Information

Received 02 May 2017 Accepted 03 July 2017 Published 01 September 2017
[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: [email protected].

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Bacterial DNA Extraction Using Individual Enzymes and Phenol/Chloroform Separation †

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Citation: Wright M, Adelskov J, Greene A. 2017. Bacterial dna extraction using individual enzymes and phenol/chloroform separation † . J. Microbiol. Biol. Educ. 18(2): doi:10.1128/jmbe.v18i2.1348

/content/jmbe/10.1128/jmbe.v18i2.1348.f1-jmbe-18-48

FIGURE 1

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

jmbe/18/2/jmbe-18-48f1_thmb.gif

jmbe/18/2/jmbe-18-48f1.gif

FIGURE 1

Agarose gel containing λHind III linear standard (A) and purified Escherichia coli 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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Bacterial DNA Extraction Using Individual Enzymes and Phenol/Chloroform Separation †

Key Concept Ranking

References & Citations

Supplemental Material

Appendix 1: Pre-laboratory questions, Appendix 2: Pre-laboratory answer key, Appendix 3: DNA extraction flowchart, Appendix 4: Agarose gel preparation for electrophoresis

Author and Article Information

Figures

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