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ERIC Number: EJ920849
Record Type: Journal
Publication Date: 2011
Pages: 4
Abstractor: As Provided
Reference Count: 11
ISBN: N/A
ISSN: ISSN-1470-8175
Purification and Analysis of Colorful Hypothetical Open Reading Frames: An Inexpensive Gateway Laboratory
DeSantis, Kara A.; Reinking, Jeffrey L.
Biochemistry and Molecular Biology Education, v39 n2 p141-144 Mar-Apr 2011
This laboratory exercise is an inquiry-based investigation developed around the core experiment where students, working alone or in groups, each purify and analyze their own prescreened colored proteins using immobilized metal affinity chromatography (IMAC). Here, we present reagents and protocols that allow 12 different proteins to be purified in parallel without specialized equipment and within a 2.5- to 3-hour undergraduate teaching laboratory. The visual feedback of purifying a colored biomolecule provides real-time emphasis of the power and simplicity of recombinant DNA technology and IMAC. As presented here in its simplest form, this laboratory occupies two laboratory periods: purification followed by SDS-PAGE analysis. As such, it can be easily inserted into the existing curriculum of a Biochemistry, Molecular Biology, Biotechnology, or even Genetics course to illustrate core concepts of central dogma and protein purification. Furthermore, the proteins in hand at the end of this 2-week module can also be used for follow-up experiments tailored to the needs, timeframe, and facilities available. (Contains 1 table and 4 figures.)
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: Journal Articles; Reports - Descriptive
Education Level: Higher Education; Postsecondary Education
Audience: N/A
Language: English
Sponsor: N/A
Authoring Institution: N/A