Overview and Learning Objectives

Overview

The polymerase chain reaction (PCR) laboratory technique is used in a variety of applications to make copies of a specific DNA sequence. This lesson describes how a PCR reaction works, what it accomplishes and its basic requirements for success. Examples of interpreting results are given for a seed storage protein (kafirins in sorghum). PCR's strengths, weaknesses and applications to plant biotechnology are explained.

Objectives

At the completion of this PCR lesson, learners will be able to:

Describe the in vivo replication processes that are converted to in vitro replication with PCR.
List the 5 chemical components of a PCR reaction and describe their roles.
List the functions of the 3 temperature cycles which are repeated during a PCR reaction.
Describe the process of observing and interpreting the results of a PCR experiment.

NOTE: The below could be standalone lesson/activities or included in this lesson

Design PCR primers that will target a specific gene or chromosome segment.
Troubleshoot problems with PCR based on the results of a PCR experiment.
List possible uses of PCR in genetic testing and in research.

Peer reviewed web lesson JNRLSE approved 2003

Development of the original lesson was supported in part by Cooperative State Research, Education, & Extension Service, U.S. Dept of Agriculture under Agreement Number 98-EATP-1-0403 administered by Cornell University and the American Distance Education Consortium (ADEC); and in part by USDA Initiative for Future Agriculture and Food Systems (IFAFS)and the Cooperative State Research, Education, & Extension Service, U.S. Dept of Agriculture under Agreement Number 00-52100-9710. A contribution of the University of Nebraska College of Agricultural Sciences and Natural Resources, Lincoln NE 68583, Journal Series 04-12. Any opinions, findings, conclusions or recommendations expressed in this publication are those of the author(s) and do not necessarily reflect the view of the U.S. Department of Agriculture.

Updates to the lesson in 2021 were supported in part by Iowa State University Libraries Open Educational Resources initiative.