lab 6a - Period 1 Lab 6a The Effect of Ampicillin and X-Gal...

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12/16/2009 Period 1 Lab 6a: The Effect of Ampicillin and X-Gal on transformed E. Coli Abstract: The objective of this experiment is to see how to transform bacteria using plasmids to make the bacteria resistant to ampicillin. From the information known, it can be inferred that if E. coli is transformed by the pBLU plasmid then only few colonies will be grown in the agar with ampicillin and the surviving E. coli will be blue if the agar also contains X-gal. In this experiment, E. coli is transformed with pBLU plasmids, which contain the ampR gene, and the lacz gene, which allows E. coli to be resistance to ampicillin and turn blue when in X-gal if successfully transformed. The results found do support the hypothesis since bacteria that were transformed were the only cells formed on the plates with LB/Amp and LB/AMP/ X-gal. Also the expected results agree with the actual results. Problem: How is bacteria transformed using plasmid to make the bacteria resistant to ampicillin? Introduction: The purpose of this lab is to show how bacteria can be transformed by plasmids. The plasmid used in this experiment is pBLU plasmid, which is used as a vector to transfer genes to bacteria. A plasmid is an extra chromosomal DNA molecule separate from the choromosomal DNA which is capable of replicating independently from the chromosomal DNA.The pBLU plasmid contains the ampR gene that codes for ampicillin resistance and the lacz gene that codes for beta-galactosidase, which breaks down galactose analog X-gal to produce a product that is blue. Only transformed cells are able to grow on the agar with ampcillin while all bacteria can grow on the agar with X-gal but only the transformed cells will produce blue colonies. The bacteria will need to become competent to induce transformation. To induce competence, CaCl 2 will be added to the bacteria and the bacteria will be heat shocked. After the bacteria become competent, the plasmid is added and the bacteria are later incubated in these types of agar. The
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This note was uploaded on 04/02/2011 for the course BIOLOGY 101 taught by Professor Martin during the Spring '08 term at Rutgers.

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lab 6a - Period 1 Lab 6a The Effect of Ampicillin and X-Gal...

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