ERIC Number: EJ758940
Record Type: Journal
Publication Date: 2004-May
Pages: 10
Abstractor: Author
ISBN: N/A
ISSN: ISSN-1470-8175
EISSN: N/A
Gene Amplification by PCR and Subcloning into a GFP-Fusion Plasmid Expression Vector as a Molecular Biology Laboratory Course
Bornhorst, Joshua A.; Deibel, Michael A.; Mulnix, Amy B.
Biochemistry and Molecular Biology Education, v32 n3 p173-182 May 2004
A novel experimental sequence for the advanced undergraduate laboratory course has been developed at Earlham College. Utilizing recent improvements in molecular techniques for a time-sensitive environment, undergraduates were able to create a chimera of a selected gene and green fluorescent protein (GFP) in a bacterial expression plasmid over the course of a single semester in a weekly 3-h laboratory period. Students designed PCR primers for amplification of the selected gene using computational DNA sequence analysis tools. During the experimental portion of the course, students amplified and ligated the target DNA into a commercially available GFP expression vector. Following transformation of the ligation product, plasmids were harvested from the resulting bacterial colonies and were analyzed by restriction digestion to confirm the creation of the chimeric GFP-DNA. This course gave students valuable experience with commonly used molecular techniques in an authentic research project. In addition, students gained experience with experimental design and execution. The techniques presented here are flexible and can be generalized for use with almost any DNA sequence and expression vector. This series also serves as an example of how faculty can adapt their ongoing research projects to the undergraduate laboratory.
Descriptors: Research Design, Science Laboratories, Molecular Biology, Genetics, Chemistry, Science Instruction, Advanced Courses, Microbiology, Teaching Methods
John Wiley & Sons, Inc. Subscription Department, 111 River Street, Hoboken, NJ 07030-5774. Tel: 800-825-7550; Tel: 201-748-6645; Fax: 201-748-6021; e-mail: subinfo@wiley.com; Web site: https://secure.interscience.wiley.com/cgi-bin/jhome/112782101
Publication Type: Guides - Classroom - Teacher; Journal Articles; Reports - Descriptive
Education Level: N/A
Audience: Teachers
Language: English
Sponsor: N/A
Authoring Institution: N/A
Grant or Contract Numbers: N/A

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