Show How to Use Restriction Mapping to Determine the Orientation

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The Answer is Restriction mapping is a technique used to determine the location of restriction sites within a DNA sequence. To determine the orientation of a restriction fragment ligated into a restriction site in a vector, you can use restriction mapping as follows:

1. First, digest the vector with the restriction enzyme used to create the original site where the fragment was ligated. This will result in two fragments, one containing the vector and the ligated fragment, and the other containing the remaining vector sequence.

2. Next, run the digested DNA on an agarose gel to separate the fragments based on size.

3. After the gel electrophoresis, visualize the DNA fragments using a staining method such as ethidium bromide. The gel will show the sizes of the fragments, allowing you to determine the orientation of the ligated fragment in the vector.

4. By comparing the sizes of the fragments with the expected sizes based on the known sequence of the vector and the ligated fragment, you can determine the orientation of the ligated fragment in the vector.

In summary, restriction mapping can be used to determine the orientation of a restriction fragment ligated into a restriction site in a vector by digesting the vector, running the fragments on an agarose gel, and comparing the sizes of the fragments to the expected sizes based on the known sequences.

Show How to Use Restriction Mapping to Determine the Orientation

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Show How to Use Restriction Mapping to Determine the Orientation

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Correct Answer:
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