Recombinant DNA Technology Explained For Beginners
Recombinant DNA technology is a series of techniques used to manipulate and isolate DNA segments of interest. In order to insert this isolate DNA into a cell for cloning, some type of vector is commonly used.
To create such a recombinant vector a bacteria containing a plasmid and a cell containing the DNA of interest is used. The plasmid from the bacteria is combined with the gene of interest that has been isolated.
This plasmid is a separate piece of circular DNA located in the bacteria and when we look closer at it we can see that it consists of complementary DNA strands just like normal DNA.
By using restriction enzymes to cut this circular DNA at two specific points, one can create two sticky ends that are complementary to the DNA of interest, allowing for easier binding of the gene of interest.
Then once the DNA strand of the gene of interest has bound to the two sticky ends of the plasmid, DNA ligase can be used to ensure proper binding.
What we are left with is a recombinant DNA plasmid which can be inserted back into a bacterial cell for cloning or into a mammalian cell for transfection.
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