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1)how many times did Pvu II cut the circular plasmid DNA?2

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1)how many times did Pvu II cut the circular plasmid DNA?2.how many times did Ava II cut the circular plasmid DNA?

3.Roughly, how big is the smallest DNA fragment in the Pvu II digest? The answer is in base pairs, but just enter a number.

4.Roughly, how big is the smallest DNA fragment in the Ava II digest? The answer is in base pairs, but just enter a number.

5.how many pieces of DNA are there in the digest with both Pvu II and Ava II?

 

6.Your objective is to create map of the pUC19 plasmid with the cut sites for both the Pvu II enzyme and the Ava II enzyme labeled, based on the fragments they create. Your solution to this puzzle, your map, should match the gel data for all four tubes. 

 

Start by creating maps for each enzyme individually.

 

Draw a circle using the bottom of a round glass or tumbler on a piece of paper. This represents the pUC19 plasmid. Now, mark the cuts made on circle - the plasmid - so that you would get a piece of DNA the same size as that produced by the digest with Pvu II.

 

Now repeat those steps but make the cuts to produce a fragment of DNA corresponding to the cuts made by Ava II.

 

Those are the single digest maps.

 

Then, in your head, rotate one of the pieces of paper on top of the other so that the other two pieces of DNA in the double digest are the same size as the two larger pieces of DNA in the double digest. If you have tracing paper, or some parchment paper or wax paper from your kitchen, you can against a window so you can see the circle on the bottom piece of paper through the top piece of paper. You may even be able to make it work with notebook paper and sharpie. Then rotate the top piece of paper until the cuts on the bottom circle in combination with the cuts on the top circle give you all the correct sized pieces of DNA.

 

When you've figured out the sites of the cuts by the enzymes, draw a new circle on a piece of paper and mark the cuts and the distances, in base pairs, between them. Label your map with the name of the plasmid (pUC19) and the total number of base pairs (2,686). Your estimated fragments should add to the total 2,686 bp.