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9th Annual Meeting 
of the 
International Association of Medical Science Educators 

July 14-19, 2005
 

Abstract Category: Methods

Poster ID: M18

A laboratory design for cloning multiple genes linked to human diseases using recombinant DAN techniques

D.P. Gardner1, T. Thal1, M. Armijo2, G. Buss3, A. Casapao2, L. Hohulin2, J. Horton2, C. Sereduk2, L. Walsh2 and Y.G. Yueh*1.  1Basic Science Department, 2 Bachelor’s of Biomedical Science, 3Master’s of Biomedical Science, Midwestern University , Glendale , AZ 85308

This integrated lecture and laboratory course is designed to offer students in biomedical and life science fields a hands-on experience and a solid background in standard molecular biology and recombinant DNA techniques.  The lecture portion explained the rationale behind differing approaches and techniques.  The laboratory part emphasized the practical application of recombinant DNA technology.  The student participants performed a variety of techniques including human genomic DNA isolation from their own buccal cells (Epicentre DNA extraction kit), PCR amplification of a selected gene linked to a known human disease (e.g. hexA/Tay-Sachs disease, SOD-1/Amyotrophic Lateral Sclerosis, HFE/Hemochromatosis and dystrophin/Duchenne Muscular Dystrophy), recombinant plasmid creation using the PCR fragment and pTarget Vector System Kit (Promega), transformation of E.coli bacteria with the recombinant plasmid, and selection, purification (Wizard Minipreps DNA Purification System by Promega) and verification of the clone using restriction enzyme digest and PCR method.  Students also performed sequence analysis using the NCBI sponsored Basic Local Alignment Search Tool (BLAST).  This course also provides extensive training in laboratory notebook as well as conventional research paper writing.  Students were evaluated at the end of 10 weeks based upon their laboratory participation, notebook writing, quizzes and research paper writing.       
 

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