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You have isolated some mitochondrial DNA (mtDNA) from walleye (Sander vitreus)
The mtDNA was digested with four restriction endonucleases Apa I, Bd II, Cla I, and Dru 1).
The fragments obtained (sizes in kilobase pairs) with each enzyme were as follows:
Apa! 11.5,3.0, 2.0
Bell! I cut only - fragment size 16.5
Cla! 11.3.52
Dra! 13.0, 3.5
You wish to map these restriction sites on the molecule, so you conduct a series of double
digests, which give the following sized fragments:
Apa VBd II 10.0,3.0, 2.0, 1.5
Apa Cla! 7.5, 4.0.2.0.1.8.1.2
Apa Dra! 9.5,3.0, 2.0, 1.5, 0.5
Bel I/Cla1 6.0.5.3, 5.2
Bel IVDra1 13.0, 3.0, 0.5
Cla VDra1 5.5, 5.2, 3.5, 2.3
Draw a lincar restriction site map for this Daphnia mtDNA that is opened at the single
Bel II restriction site. Show your working, including alternative positions of restriction sites in
the preliminary maps
A polymorphism is now found using Dra I such that the new fragment pattern for this
enzyme becomes: 8.0.5.0.3.5. What has happened?
You conduct two more double digests in order to map this variable site, giving results as
follows:
Cla I Dual 5.2, 5.0, 3.5, 2.3, 0.5
Apa Dra! 5.0.4.53.0, 2.0, 1.5, 0.5
Map the variable site and mark it with an
You examine another individual with Dra I revealing the following fragment results: 9.0,
5.0, 3.5. You think that this is a bit odd. So you conduct a few more digests, getting the
following results:
Apal 11.5, 4.0, 20
Cla! 11.3.6.2
Where is this phenomenon affecting the mtDNA molecule? Indicate the portion of the
molecule that is affected. What fragments would you expect to result from a Cla VDra I double
digest on this individual?
Draw a picture of a gel showing all of the fragments obtained in the above digests.
Total points - 50