Restriction Enzyme Practice - Jennifer Kaltenbach | Library

Use the following sequences of DNA and use the particular restriction endonuclease to cut the DNA. In each case you will need to search for recognition sites on both strands of DNA, forward and backward for the restriction sequence. The entire sequence of the restriction enzyme (all the letters) and in that exact order are both needed to make a cut. There are four parts to your assignment for each of the 6 practice problems below. When you find the recognition sequence - (1) draw a box around it precisely. Now, use a colored pen to (2) mark the cut(s) on the DNA; be sure to exactly follow the pattern shown on the previous page for each restriction enzyme. Label each as blunt cut or sticky end cut (3). Finally, count up the number of cuts (4) you found and write it to the right of the DNA with blunt or sticky. An answer of no cuts is just as good as many cuts! Remember to go in both directions (forward and backward) on both strands.

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Use ThaI to cut the following DNA:

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What type of cut did you make?

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How many cuts did you make?

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Use BamHI to cut the following DNA:

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What type of cut did you make?

1

How many cuts did you make?

1

Use SmaI to cut the following DNA:

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What type of cut did you make?

1

How many cuts did you make?

1

Use TaqI to cut the following DNA:

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What type of cut did you make?

1

How many cuts did you make?

1

Use TaqI to cut the following DNA:

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What type of cut did you make?

1

How many cuts did you make?

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Use BOTH BalI and BglII to cut this DNA:

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What type of cut did you make?

1