 OBJECTIVES:Molecular Biology
REVIEW OF A BACTERIAL TRANSFORMATION KIT
Bacterial Transformation: Review of the BioRad kit.
The PGLO System for Bacterial Transformation
Catalogue Number 166-0003-EDU
Bio Rad corporation (1-800-424-6723)
As an alternative to a basic bacterial transformation lab sold by many biological supply companies, we had the opportunity to work with a "variation on a familiar theme". The pGLO kit offered by a company called BIO - RAD is an interesting change of pace! Instead of just noticing transformation by a color change, transformed cells actually glow when they are exposed to ultraviolet light.
This particular kit is purchased with enough materials for 6 student stations. The transformation involves moving a gene from a bioluminescent jellyfish which makes a protein called Green Flourescent Protein (GFP). When introduced into E. coli, the colonies will flouresce a brilliant green/yellow. Not included with the kit but necessary is an inexpensive hand help UV light source. We used the common model sold for use with minerals. It worked beautifully. The results are indeed striking. Another unusual feature is a much extended appendix of additional exercises for students. The directions generally are very clear and easy to follow .
No kit is perfect, so there are a few things about the preparation procedures that you may want to revise
1. The directions say that starter cells are good after 24-36 hours. We found that 36 hours makes a more satisfactory starter plate.
2. I would also recommend in following the directions to modify the procedure for preparing the nutrient LB agar. The directions say to heat the water and then add the packet of agar to the hot water and continue to microwave. This resulted in a really messy set of lumps. Instead we found that if you add about 300 of the needed 500 ml room temperature water to the flask, and then add the agar powder to the cold water. You can get a very nice mixing just by swirling. Then you can microwave this and it goes quickly and without a hitch. When the agar is properly dissolved, we added the remaining 200ml of water to the hot agar. It was a snap that way. You can also use the hotplate/stirrer method.
We noted that there is an upcoming kit that can be used to then extract the new glo protein from the transformed cells. This might prove a very interesting follow up. While we have not tried this, if anyone does, please send in the a review of it.
(**NOTE: There is no spreadsheet for Lab 6.)
6/28/97 Judy Owens
St. Pius X High School, PA
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Laboratory Section
AP Bio Index