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ERIC Number: EJ1234161
Record Type: Journal
Publication Date: 2019
Pages: 6
Abstractor: As Provided
ISBN: N/A
ISSN: ISSN-1470-8175
EISSN: N/A
Available Date: N/A
Plasmid Production and Purification: An Integrated Experiment-Based Biochemistry and Biotechnology Laboratory Course
Santos, Tiago; Pereira, Patrícia; Queiroz, João A.; Cruz, Carla; Sousa, Fani
Biochemistry and Molecular Biology Education, v47 n6 p638-643 Nov-Dec 2019
This laboratory experiment describes the production and purification of plasmid DNA for undergraduate biochemistry and biotechnology courses. This experiment performed in a one-week period includes the protocols for plasmid pVAX1-LacZ production in "Escherichia coli" DH5[alpha] cells and subsequent purification of supercoiled pVAX1-LacZ. Firstly, the students use a growth medium that favors the replication of the plasmid resulting in a higher plasmid production during exponential growth. Afterwards, alkaline lysis is done to disrupt the bacterial cells and recover pVAX1-LacZ plasmid. Lastly, they perform the purification of pVAX1-LacZ supercoiled isoform by L-histidine chromatography, followed by agarose gel electrophoresis to characterize the separation of supercoiled isoform from contaminants. The proposed experiment provides an opportunity for students to acquire these skills that are routinely used in biochemistry and biotechnology laboratories.
Wiley-Blackwell. 350 Main Street, Malden, MA 02148. Tel: 800-835-6770; Tel: 781-388-8598; Fax: 781-388-8232; e-mail: cs-journals@wiley.com; Web site: http://www.wiley.com/WileyCDA
Publication Type: Journal Articles; Reports - Descriptive
Education Level: Higher Education; Postsecondary Education
Audience: N/A
Language: English
Sponsor: N/A
Authoring Institution: N/A
Grant or Contract Numbers: N/A
Author Affiliations: N/A