Question

Genetic/Biology question-

Restriction Mapping and Creating and Designing Recombinant DNA.

I have a human DNA sequence consisting of 868 bp,

of which 3 specific restriction enzymes made cuts.

I was then given a plasmid DNA sequence of 2220 bp and I'm asked to ligate the DNA into a plasmid DNA vector.

The plasmid DNA sequence also had one of the same restriction enzymes as the human sequence...

ex: the cut is made on the human sequence at bp 101, and the same restriction enzyme cuts the plasmid sequence at bp 460.

My question- when ligating and creating a vector do I add both of the sequences together after the matching EcoRI restriction enzyme cuts each?

Is ligating essentially adding the two together at the cut site of each?

How would I draw this?

I'm super confused

Answer 1

Yes you need to make both of the sequences together. The ligation mixture which you will prepare will contain -

1. Cut plasmid

2. Cut foreign gene

3. DNA ligase enzyme

4. Buffer

5. Water (to make up the final volume)

You need an enzyme which cut two times on the human gene and one in the plasmid.

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