DNA Amplification by PCR and Analysis by Gel Electrophoresis-An Undergraduate Biochemistry Experiment

TCE

The Chemical Educator

ISSN: 1430-4171 (electronic version)

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Abstract Volume 9 Issue 2 (2004) pp 97-101

DNA Amplification by PCR and Analysis by Gel Electrophoresis—An Undergraduate Biochemistry Experiment

Beng Guat Ooi

Department of Chemistry, Middle Tennessee State University, Murfreesboro, TN 37132, bgooi@mtsu.edu
Received July 2, 2003. Accepted February 7, 2004.

Published online: 16 March 2004

Abstract. The purpose of this laboratory experiment is to introduce students to the polymerase chain reaction (PCR); to demonstrate how the PCR can be used to amplify a specific region of the genome using small amounts of DNA extracted from yeasts; and how the amplification produced sufficient DNA for further studies such as DNA sequencing, restriction enzyme analysis, and even for cloning purposes. Students will acquire skills in the extraction of genomic DNA from a yeast-cell culture, the determination of cell density, and the quantization of DNA using a spectrophotometer. They will also learn how to estimate the size of DNA fragments using agarose gel electrophoresis by comparing it to DNA calibration standards.

Key Words: Laboratories and Demonstrations; biochemistry; PCR; electrophoresis; DNA analysis

(*) Corresponding author. (E-mail: bgooi@mtsu.edu)

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