P838: Teaching “Beyond the Kit” in biochemistry lab

Author: Dana Emmert, Warren Wilson College, USA

Co-Author:

Date: 8/6/14

Time: 2:45 PM3:05 PM

Room: LOH 174

Related Symposium: S54

At Warren Wilson College we are moving “beyond the kit” in our biochemistry lab. We are extending the resources available from current educational kits. For example, using the pGLOTM Bacterial Transformation Kit (Bio-Rad), students would transform pGLO plasmid DNA into E. coli, determine transformation efficiency, and identify GFP using UV light. We extended this to include the purification of additional pGLO DNA and subsequent generation of a restriction map using a combination of restriction digests. The students then utilized online databases to compare the pGLO GFP DNA and amino acid sequences to that of the original organism, A. victoria. The lab also delved into the possible effect of these mutations on protein structure and solubility by viewing the 3D structure of GFP. This experimental sequence was followed by the expression and purification of GFP using the GFP Purification Kit (Bio-Rad). Students are currently extending this to include detecting the concentration of purified protein and SDS-PAGE. Students will perform different protein concentration assays and staining methods to compare sensitivity. These additional lab experiments have allowed the generation of a multi-week GFP based “workshop”. The students learned not only hands-on lab procedures, but also how to utilize online electronic resources. We hope to further expand on this area by introducing a similar multi-week independent project, allowing the students to utilize the skills acquired during the GFP “workshop” to characterize a protein of their choice (selected from a short list). This portion will be expanded to include activity studies of the selected proteins.