Application of the Polymerase Chain Reaction (PCR) Technique

Application of the Polymerase Chain Reaction (PCR) Technique



This CREATE module, composed of 2 papers, will be used to reinforce the application of the Polymerase Chain Reaction (PCR) technique. Before the module is introduced, students will have just finished DNA isolation from their own cheek cells, PCR amplification using primers specific to the PV92 region of chromosome 16, gel electrophoresis, and data analysis of their samples. After this module, the students will revisit analysis of their own data and results.


Applicable for Courses:

Molecular Biotechnology

Educational Level:

Roadmap Objectives:

    • Article: Comas et al. (2001) Recent Insertion of an Alu Element Within Polymorphism Human-Specific Alu Insertion. Mol. Biol. Evol. 18(1):85-88.
    • Content area/major concepts: I chose this paper because it uses the same region of the genome (PV92) that students amplified in the lab. It extends the students inquiry into this region and introduces DNA sequencing.

      molecular evolution, Mendellian genetics, molecular genetics, transposons, bioinformatics, DNA polymorphism, human migration patterns.
    • Methods or technology used to obtain data: PCR, sequencing, genomic DNA extraction, DNA extraction from agarose gels, PCR and sequencing primer design.
    • How the CREATE strategy was used:
    • Biggest teaching challenge: Students making the connection between banding patterns on gels and Mendellian genetics. Also, fig. 2b annotation may be confusing
    • Article: Agostini et al. (1997) Asian genotypes of JC virus in Native Americans and in a Pacific Island population: Markers of viral evolution and human migration. Proc. Natl. Acad. Sci. USA vol. 94, pp. 14542-14546
    • Content area/major concepts: I chose this as the next paper in the module because it continues to reinforce ideas about application of PCR and its use in the study of human migration. From the previous paper (although not sequential), it extends the study of human migration by using PCR of a human virus, and introduces an additional type of DNA sequencing.

      Neuron anatomy, neurodegenerative disease, virology, virion structure, PCR and sequencing primer design, genetics of JCV, human migration, evolution, protein structure, statistics, genotyping, viral evolution, scientific collaboration.
    • Methods or technology used to obtain data: Collection of clinical samples, collaboration across states in the U.S., DNA isolations, PCR, cycle sequencing, statistics analysis software.
    • How the CREATE strategy was used: I will ask students to consider both papers and suggest experiments to reinforce the Bering Land Bridge conclusion of both papers. I will then have students to re-visit the class data on Alu insertion in the PV92 region of chromosome 16 and ask if they want to do any follow up experiments of their own work.
    • Biggest teaching challenge: Making connections between data and human migration patterns.

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