Question

- What information do you obtain from the double digest XhoI/AlwNI? What is your expectation if the plasmid is a reclosed vector without an insert? What is your expectation if your plasmid has an insert in the correct orientation (how many bands, what are their sizes)? - What is your expectation if your plasmid has an insert in the incorrect orientation (how many bands, what are their sizes)?

Answer 1

Plasmids are used as vectors in recombination technology to transfer genes. Plasmids or the genes can be double digested by two restriction endonucleases simultaneously.

What are restriction endonucleases?

Restriction endonucleases are the enzymes that identify the precise DNA sequence to cleave them from the rest of the sequence.

If a plasmid is reclosed vector without an insert then only one band of size 2964 bp will be seen. A plasmid that has correct orientation will have two bands of sizes 1570 and 2100 bp.

If the plasmid has a sequence in incorrect orientation then will also result in two bands but of different sizes like 2740 bp and 940 bp.

Therefore, plasmids can be used as vectors.

Learn more about plasmids here:

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