Answers_to_Post_lab - D Control to show that many live E...

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Question 1. Briefly described below are various functions of each of the following steps in Cohen, Chang, Boyer, Helling experiment: 1 Competent E-coli cells -L 2 Electrophoresis of the Restriction Mixture -A 3 Electrophoresis of the Ligation Mixture -C 4 Incubation of Bacteria and Plasmids in Cold CaCl 2 -B 5 Heat Shocking Treatment -E 6 Recovery After Heat Shocking -F 7 Plates Without Antibiotics -D 8 Plates with Both Antibiotics -K 9 Miniprep Gel -J 10 Ethidium Bromide -G 11 Wild type E-coli control plates during selection -H 12 Standard curve base don migration of Lambda phage fragments -I A. Verify that restriction with restriction enzymes was successful or estimate the size of the plasmid fragments. B. Neutralize negative charges on cell membrane and plasmid DNA. C. Verify that ligation was successful.
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Unformatted text preview: D. Control to show that many live E. coli cells were plated and can grow in the absence of antibiotics. E. Facilitates the movement of plasmids into the cells. F. Allows plasmids to replicate,which amplifies their genes. G. Binds to DNA and fluoresceses in UV radiation so DNA bands can be seen on a gel. H. These ascertain the effectiveness of the antibiotics used and establish that they are not resistant to these antibiotics. I. Helps determine the size of Ampr and Kanr plasmids. J. Determine the types and composition of plasmids contained by a colony (clone) of cells growing on a double antibiotic plate. K. Select for cells that have resistance genes for both antibiotics. L. Able to take in more readily and express foreign DNA....
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This note was uploaded on 05/18/2008 for the course BIO G 104 taught by Professor Chen,k.c during the Spring '06 term at Cornell.

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