Look carefully at the differences between the four plasmids. We will use the pat

Question

Look carefully at the differences between the four plasmids. We will use the pattern that results
from reaction with restriction enzymes to identify our plasmid. As an example, of what we may
see, digests by either EcoRI or AvaI will simply give you a piece that is the size of the entire
plasmid because there is only one cutsite for each of them. However, a double digest with
both EcoRI and AvaI will allow you to distinguish pAB125 from the others. Plasmid pAB125 will
give a large fragment ~4000 bp in length and a small fragment ~300 bp in length. In contrast,
the smaller fragment resulting from a double digest with the other three plasmids would be a
clearly distinguishable ~1500 bp in length. Answer these questions:
1. Which of the four plasmids will be clearly identifiable from a RSA1 digest?
2. What digest experiment would clearly identify pAB122?

Explanation / Answer

1. If we look closely the vector map of all the plasmid we will find that in the case of plasmid pAB124 will give three fragments on digestion with RsaI whereas pAB123, pAB122 and pAB125 will give two digested fragments.

2. Digestion with HincIII only linearizes the pAB122 vector whereas when other vectors are digested with HincIII they will give 2 fragments each.

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