Bio 210-2 Lab4 - Jason Kamin Bio 210-2 3/10/08 Exercise 4:...

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Jason Kamin Bio 210-2 3/10/08 Exercise 4: Two-Hybrid Assay Introduction In this lab we are looking to identify protein-protein interactions through a two- hybrid assay. These interactions will eventually transcribe for some process or event inside a cell that can control the fate of the cell or tell the cell what to do. When combining and plating yeast with different proteins, we will be able to tell whether or not these proteins interacted and activated the transcription of a reporter gene by whether or not the yeast grew on the various media. From this we will be able to make conclusions about certain test proteins used in the experiment and the interactions in which they take part. Materials and Methods Part I: Yeast Transformation: In this part of the lab we were to test out the bait and prey plasmids that were marked at out bench. After receiving 100 ul of yeast cells and adding 10 ul of bait, prey and carrier DNA, respectively, to the cells, we mixed by vortexing for 15 seconds. Various other solutions were then added to this mixture, including polyethylene glycol/lithium acetate, DMSO and TRIS/EDTA (TE), while intermittently vortexing, incubating, heating, chilling and centrifuging the mixture. Finally, 200 ul of the yeast mixture was added to the center of two Trp minus/Leu minus
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This note was uploaded on 02/16/2012 for the course BIOL_SCI 210-2 taught by Professor Somebody during the Winter '09 term at Northwestern.

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Bio 210-2 Lab4 - Jason Kamin Bio 210-2 3/10/08 Exercise 4:...

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