1887

Using PCR to Target Misconceptions about Gene Expression

    Authors: Leslie K. Wright1,*, Dina L. Newman1
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    Affiliations: 1: Thomas H. Gosnell School of Life Sciences, Rochester Institute of Technology, Rochester, NY 14623
    AUTHOR AND ARTICLE INFORMATION AUTHOR AND ARTICLE INFORMATION
    • Published 06 May 2013
    • Supplemental materials available at http://jmbe.asm.org
    • *Corresponding author. Mailing address: School of Life Sciences, Rochester Institute of Technology, 85 Lomb Memorial Drive, Rochester, NY 14623-5608. Phone/Fax: 585-475-4449. E-mail: lkwsi@rit.edu.
    • ©2013 Author(s). Published by the American Society for Microbiology.
    Source: J. Microbiol. Biol. Educ. May 2013 vol. 14 no. 1 93-100. doi:10.1128/jmbe.v14i1.539
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    Abstract:

    We present a PCR-based laboratory exercise that can be used with first- or second-year biology students to help overcome common misconceptions about gene expression. Biology students typically do not have a clear understanding of the difference between genes (DNA) and gene expression (mRNA/protein) and often believe that genes exist in an organism or cell only when they are expressed. This laboratory exercise allows students to carry out a PCR-based experiment designed to challenge their misunderstanding of the difference between genes and gene expression. Students first transform with an inducible GFP gene containing plasmid and observe induced and un-induced colonies. The following exercise creates cognitive dissonance when actual PCR results contradict their initial (incorrect) predictions of the presence of the GFP gene in transformed cells. Field testing of this laboratory exercise resulted in learning gains on both knowledge and application questions on concepts related to genes and gene expression.

Key Concept Ranking

Agarose Gel Electrophoresis
0.42883047
Amino Acid Synthesis
0.4255062
0.42883047

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/content/journal/jmbe/10.1128/jmbe.v14i1.539
2013-05-06
2017-12-14

Abstract:

We present a PCR-based laboratory exercise that can be used with first- or second-year biology students to help overcome common misconceptions about gene expression. Biology students typically do not have a clear understanding of the difference between genes (DNA) and gene expression (mRNA/protein) and often believe that genes exist in an organism or cell only when they are expressed. This laboratory exercise allows students to carry out a PCR-based experiment designed to challenge their misunderstanding of the difference between genes and gene expression. Students first transform with an inducible GFP gene containing plasmid and observe induced and un-induced colonies. The following exercise creates cognitive dissonance when actual PCR results contradict their initial (incorrect) predictions of the presence of the GFP gene in transformed cells. Field testing of this laboratory exercise resulted in learning gains on both knowledge and application questions on concepts related to genes and gene expression.

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Figures

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FIGURE 1

Typical depiction of the Central Dogma concept.

Source: J. Microbiol. Biol. Educ. May 2013 vol. 14 no. 1 93-100. doi:10.1128/jmbe.v14i1.539
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Image of FIGURE 2

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FIGURE 2

Sophomore-level biology students do not demonstrate a clear understanding of PCR or the difference between DNA replication and expression. Students from a second-year Molecular Biology course (n = 49) predicted the results of their PCR experiment with the open-ended question shown in Appendix 3 (Pre-lab assessment). Categories of student responses are as follows: 1) Gene (DNA) is present whether or not it is being expressed; 2) Amount of gene (DNA) present is proportional to expression level of gene; 3) Gene (DNA) is only present when it is being expressed; 4) Presence of arabinose allows the GFP gene to get added to the E. coli genome; 5) Unable to follow logic.

Source: J. Microbiol. Biol. Educ. May 2013 vol. 14 no. 1 93-100. doi:10.1128/jmbe.v14i1.539
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Image of FIGURE 3

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FIGURE 3

PCR amplification of the GFP gene from pGLO Examples of three student gels demonstrating GFP gene amplification (white arrows at 714 bp) in samples labeled G+ (DNA from induced pGLO ) and G– (DNA from uninduced pGLO ) but not W (DNA from wild-type DNA ladders (L) are included on all gels. White arrows indicate the 714 bp PCR product. Black arrows indicate primer-dimer product, which varies in intensity depending on conditions.

Source: J. Microbiol. Biol. Educ. May 2013 vol. 14 no. 1 93-100. doi:10.1128/jmbe.v14i1.539
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