Bio 210-2 Lab 3 - Jason Kamin Bio 210-2 3/6/08 Exercise 3:...

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Jason Kamin Bio 210-2 3/6/08 Exercise 3: Transformation and Restriction Mapping Introduction In this lab the goal was to insert foreign DNA, in the form of a plasmid, into host cells by bacterial transformation. In this case the bacterium that was worked with was E. coli. At first the E. coli was screened for plasmid-carried drug resistance markers by testing whether or not they would grow on various drug-containing media, with separate media containing the drugs ampicillin (amp) and kanamycin (kan). There was also a control media containing no drug. Next, the plasmid DNA was removed from the bacteria and it was analyzed for its size by using restriction enzyme mapping and gel electrophoresis. From the data that was received we were able to create a restriction map of the plasmid and determine the locations of the cut sites. Materials and Methods Part I: Transformation: In this part we were to perform the transformation of getting the foreign DNA plasmid into the E. coli bacteria and plating these cells. After receiving the microcentrifuge tube containing E. coli in growth medium we proceeded to do a number of centrifuging steps, subsequently adding certain solutions like transformation buffer, DMSO, plasmid, and growth medium. After this process of centrifugation, chilling, thawing, and incubating, the cells were spread onto three petri dishes, one a control, one with amp and one with kan, and then allowed to incubate for 1-2 days or until colonies appeared. Part IIA: Overnight Culture:
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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 Lab 3 - Jason Kamin Bio 210-2 3/6/08 Exercise 3:...

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