1887

Designing PCR Primers Painlessly

    Authors: Morgan Feeney1, Kevin Murphy1, Jane Lopilato1,*
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    Affiliations: 1: Biology Department, Simmons College, Boston, MA 02115
    AUTHOR AND ARTICLE INFORMATION AUTHOR AND ARTICLE INFORMATION
    • Published 01 May 2014
    • Supplemental materials available at http://jmbe.asm.org
    • *Corresponding author. Mailing address: Biology Department, Simmons College, 300 The Fenway, Boston, MA 02115. Phone: 617-521-2661. Fax: 617-521-3086. E-mail: jane.lopilato@simmons.edu.
    • ©2014 Author(s). Published by the American Society for Microbiology.
    Source: J. Microbiol. Biol. Educ. May 2014 vol. 15 no. 1 28-29. doi:10.1128/jmbe.v15i1.634
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    Abstract:

    Students design primers using web-based tools along with a printed sequence of a prokaryotic gene showing both DNA strands and translation of the coding strand into amino acids. The melting temperature of each primer is determined and the pair closest in temperature can be used to amplify and clone a gene into a vector for a one-semester laboratory exercise. By working through this primer design exercise, students see first hand what is involved in the process of amplifying DNA.

Key Concept Ranking

DNA Synthesis
0.6929444
DNA
0.4935737
Escherichia coli
0.4935737
DNA
0.4935737
Escherichia coli
0.4935737
DNA
0.4935737
Stop Codon
0.45169035
0.6929444

References & Citations

1. Guzman L-M, Belin D, Carson MJ, Beckwith J 1995 Tight regulation, modulation, and high-level expression by vectors containing the arabinose PBAD promoter J. Bacteriol 177 4121 4130 7608087 177145
2. Klein JR, Gulsvig T 2012 Using bioinformatics to develop and test hypotheses: E. coli-specific virulence determinants J. Microbiol. Biol. Educ 13 161 169 10.1128/jmbe.v13i2.451 23653804 3577336 http://dx.doi.org/10.1128/jmbe.v13i2.451
3. May BJ 2013 Engaging students in a bioinformatics activity to introduce gene structure and function J Microbiol Biol Educ 14 107 109 10.1128/jmbe.v14i1.496 23858361 3706140 http://dx.doi.org/10.1128/jmbe.v14i1.496
4. Simons RW, Houman F, Kleckner N 1987 Improved single and multicopy lac- based cloning vectors for protein and operon fusions Gene 53 85 96 10.1016/0378-1119(87)90095-3 http://dx.doi.org/10.1016/0378-1119(87)90095-3
5. Wright LK, Newman DL 2013 Using PCR to target misconceptions about gene expression J Microbiol Biol Educ 14 93 100 10.1128/jmbe.v14i1.539 23858358 3706170 http://dx.doi.org/10.1128/jmbe.v14i1.539
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/content/journal/jmbe/10.1128/jmbe.v15i1.634
2014-05-01
2017-09-21

Abstract:

Students design primers using web-based tools along with a printed sequence of a prokaryotic gene showing both DNA strands and translation of the coding strand into amino acids. The melting temperature of each primer is determined and the pair closest in temperature can be used to amplify and clone a gene into a vector for a one-semester laboratory exercise. By working through this primer design exercise, students see first hand what is involved in the process of amplifying DNA.

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