F08 LS1a Pset 6keyfinalst-1

F08 LS1a Pset 6keyfinalst-1 - Name: TF Name: LS1a Fall 08...

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Name: TF Name: 1 LS1a Fall 08 Problem Set #6 Due Monday 11/10 at noon in your TF’s drop box on the 2 nd floor of the Science Center (100 points total) 1. (44 points) The Polymerase Chain Reaction (PCR) is a useful tool to amplify specific DNA sequences. A gene sequence within a long stretch of DNA is shown below diagrammatically. GCATGG CCAATG 5' 3' 3' 5' GGATCC a) Write the DNA base sequences corresponding to the “light blue” and the “light green” regions on the bottom strand. Be sure to designate 5’ and 3’ ends of your sequences. (4 points) “light blue”: 5’-CCATGC-3’ “light green”: 5’-CATTGG-3’ b) Write the sequences of the two primers needed to amplify the gene beginning with the blue sequence and ending with the green sequence. Be sure to designate 5’ and 3’ ends. (4 points) “dark blue”: 5’-GCATGG-3’ and “light green”: 5’-CATTGG-3’ For the following questions (c-e) include the sequence of the blue and green regions (both dark and light) in your representations and designate 5’ and 3’ ends. For instance the top strand should be written as: GCATGG CCAATG 3' 5' c) Draw the hybridized DNA that would form if you took only the top strand of DNA and mixed it with the two primers from part b. (4 points) GCATGG CCAATG 3' 5' 3' -GGTTAC- 5' Gene Sequence
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Name: TF Name: 2 d) Draw the hybridized DNA that would form if you took only the bottom strand of DNA and mixed it with the two primers from part b. (4 points) CGTACC GGTTAC 5' 3' 5' -GCATGG- 3 e) Would you get the same products for the first three cycles if you used only one DNA primer instead of two primers in your PCR reaction? Briefly explain. (6 points) No, you would not get the same products as in part (d). You would only get a single new product from the primer hybridizing to the template in all three cycles. [Alternate answer: Yes, if you used only one primer you would get the same product in each of the first three cycles since the primer can only hybridize to the original template strand. When you have two primers as in part (d) they can hybridize not only to the original template
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F08 LS1a Pset 6keyfinalst-1 - Name: TF Name: LS1a Fall 08...

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