Who Scared the Cat? A Molecular Crime Scene Investigation Laboratory Exercise †
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Authors:
Laura E. Ott1,*,
Susan D. Carson2
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Published 02 December 2016
- ©2016 Author(s). Published by the American Society for Microbiology.
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[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.
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Supplemental materials available athttp://asmscience.org/jmbe - *Corresponding author. Mailing address: University of Maryland, Baltimore County, College of Natural and Mathematical Sciences, 1000 Hilltop Circle, Baltimore, MD 21250. Phone: 410-455-8089. Fax: 410-455-5831. E-mail: [email protected].
Abstract:
This introductory laboratory exercise gives first-year life science majors or nonmajors an opportunity to gain knowledge and experience in basic bioinformatics and molecular biology laboratory techniques and analysis in the context of a mock crime scene investigation. In this laboratory, students determine if a human (Lady) or dog (Kona) committed the fictional crime of scaring a cat. Students begin by performing in silico PCR using provided dog- and human-specific PCR primers to determine the sequences to be amplified and predict PCR amplicon sizes. They then BLAST (Basic Local Alignment Search Tool) the in silico PCR results to confirm that the PCR primers are designed to amplify genomic fragments of the cardiac actin gene in both dogs and humans. Finally, they use DNA quantification techniques, PCR, and agarose gel electrophoresis to identify the culprit and they confirm results by analyzing Sanger sequencing. Student learning gains were demonstrated by successful execution of the lab and by analysis and interpretation of data in the completion of laboratory reports. The student learning gains were also demonstrated by increased performance on a post-laboratory assessment compared to the pre-assessment. A post-activity assessment also revealed that students perceived gains in the skills and conceptual knowledge associated with the student learning outcomes. Finally, assessment of this introductory molecular biology and bio-informatics activity reveals that it allows first-year students to develop higher-order data analysis and interpretation skills.
References & Citations
Supplemental Material
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Appendix 1: Lecture materials
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MyBook is a cheap paperback edition of the original book and will be sold at uniform, low price.
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Appendix 2: Instructor materials
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Appendix 3: Student handout
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MyBook is a cheap paperback edition of the original book and will be sold at uniform, low price.
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Appendix 4: Sequencing results
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MyBook is a cheap paperback edition of the original book and will be sold at uniform, low price.
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PDF
9.20 MB
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PDF
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Appendix 5: Lab report guidelines, rubric, and sample lab report
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MyBook is a cheap paperback edition of the original book and will be sold at uniform, low price.
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PDF
9.20 MB
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PDF
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Appendix 6: Pre- and post-quiz with answer key
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MyBook is a cheap paperback edition of the original book and will be sold at uniform, low price.
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PDF
9.20 MB
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Appendix 7: Other possible activity modifications
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MyBook is a cheap paperback edition of the original book and will be sold at uniform, low price.
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PDF
9.20 MB
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PDF
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Abstract:
This introductory laboratory exercise gives first-year life science majors or nonmajors an opportunity to gain knowledge and experience in basic bioinformatics and molecular biology laboratory techniques and analysis in the context of a mock crime scene investigation. In this laboratory, students determine if a human (Lady) or dog (Kona) committed the fictional crime of scaring a cat. Students begin by performing in silico PCR using provided dog- and human-specific PCR primers to determine the sequences to be amplified and predict PCR amplicon sizes. They then BLAST (Basic Local Alignment Search Tool) the in silico PCR results to confirm that the PCR primers are designed to amplify genomic fragments of the cardiac actin gene in both dogs and humans. Finally, they use DNA quantification techniques, PCR, and agarose gel electrophoresis to identify the culprit and they confirm results by analyzing Sanger sequencing. Student learning gains were demonstrated by successful execution of the lab and by analysis and interpretation of data in the completion of laboratory reports. The student learning gains were also demonstrated by increased performance on a post-laboratory assessment compared to the pre-assessment. A post-activity assessment also revealed that students perceived gains in the skills and conceptual knowledge associated with the student learning outcomes. Finally, assessment of this introductory molecular biology and bio-informatics activity reveals that it allows first-year students to develop higher-order data analysis and interpretation skills.

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Author and Article Information
-
Published 02 December 2016
- ©2016 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 athttp://asmscience.org/jmbe - *Corresponding author. Mailing address: University of Maryland, Baltimore County, College of Natural and Mathematical Sciences, 1000 Hilltop Circle, Baltimore, MD 21250. Phone: 410-455-8089. Fax: 410-455-5831. E-mail: [email protected].
Figures
Flowchart for the crime scene laboratory activity. These are the five parts to this crime scene lab, with the approximate timing of each. It is recommended that the activity span three 3-hour laboratory sessions, with parts 1, 2, and 3 performed in session 1, part 4 in session 2, and part 5 in session 3. PCR = polymerase chain reaction; DNA = deoxyribonucleic acid; BLAST = basic local alignment search tool.

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FIGURE 1
Flowchart for the crime scene laboratory activity. These are the five parts to this crime scene lab, with the approximate timing of each. It is recommended that the activity span three 3-hour laboratory sessions, with parts 1, 2, and 3 performed in session 1, part 4 in session 2, and part 5 in session 3. PCR = polymerase chain reaction; DNA = deoxyribonucleic acid; BLAST = basic local alignment search tool.
Representative agarose gel electrophoresis results from a student PCR. The sizes (base pairs; bp) of the low DNA mass ladder are depicted on the left. The samples were loaded in the following order (left to right): low DNA mass ladder (ladder), Kona control DNA in dog PCR master mix (MM), Lady control DNA in human MM, crime scene DNA in dog MM, crime scene DNA in human MM. Expected dog and human PCR amplicons are observed in the Kona control (275 bp) and Lady control (397 bp) samples, respectively. This gel depicts Lady (human) as the culprit, as there is an approximate 400 bp band in the crime scene sample in human MM. PCR = polymerase chain reaction; DNA = deoxyribonucleic acid; MM = master mix.

Click to view
FIGURE 2
Representative agarose gel electrophoresis results from a student PCR. The sizes (base pairs; bp) of the low DNA mass ladder are depicted on the left. The samples were loaded in the following order (left to right): low DNA mass ladder (ladder), Kona control DNA in dog PCR master mix (MM), Lady control DNA in human MM, crime scene DNA in dog MM, crime scene DNA in human MM. Expected dog and human PCR amplicons are observed in the Kona control (275 bp) and Lady control (397 bp) samples, respectively. This gel depicts Lady (human) as the culprit, as there is an approximate 400 bp band in the crime scene sample in human MM. PCR = polymerase chain reaction; DNA = deoxyribonucleic acid; MM = master mix.
Pre- and post-quiz results from the fall 2014 and spring 2015 semesters. Median scores (with upper and lower limits) are displayed, with 16 students completing the pre- and post-quizzes in fall 2014 and seven students completing the quizzes in spring 2015. Data was analyzed with a Wilcoxon signed-rank test (SPSS Statistics, version 22), with p-values displayed.

Click to view
FIGURE 3
Pre- and post-quiz results from the fall 2014 and spring 2015 semesters. Median scores (with upper and lower limits) are displayed, with 16 students completing the pre- and post-quizzes in fall 2014 and seven students completing the quizzes in spring 2015. Data was analyzed with a Wilcoxon signed-rank test (SPSS Statistics, version 22), with p-values displayed.