215-Lab6A-Bacterial_Tsf

215-Lab6A-Bacterial_Tsf - Lab 6A Bacterial Transformations...

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Lab 6A – Bacterial Transformations 6A-1 Lab 6A: Bacterial Transformations Background: In the last lab you purified the plasmid DNA from yeast. The yield of DNA from these preps out of yeast is very small and there is not sufficient quantity to perform the restriction digest and PCR mapping assays, sequencing of the mutant chimera gene, or transformation into another yeast strain to perform the transcriptional repression assays. We therefore need to obtain larger quantities of the plasmid. To do this you will transform the plasmid into “competent” bacteria cells. The plasmid will be highly amplified in the bacteria culture and the yields of plasmid DNA from bacteria are significantly better than from yeast. 1. Take two tubes of bacterial competent cells from the freezer storage box and place them in an ice bucket to thaw. Note: It is very important that the cells are on ice and that you do not heat the cells up with your hand or by leaving them at room temperature! Sometimes the cells are frozen at the top of the tube.
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This note was uploaded on 10/23/2011 for the course BIO 201 taught by Professor Frenkel/cervantes during the Spring '10 term at Rutgers.

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215-Lab6A-Bacterial_Tsf - Lab 6A Bacterial Transformations...

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