Pre-lab

Outline a basic technique used for gene transfer involving plasmids, a host cell (bacterium, yeast or other cell), restriction enzymes (endonucleases) and DNA ligase.

Gene transfer involving plasmids are usually used to alter the circular DNA molecules called plasmids that are usually found in plasmids. 

Stage 1: Obtaining a gene to transfer

-Restriction enzymes are put in the same container as the DNA strands

-During this stage, restriction enzymes are used to cut the targeted gene that is to be transferred from the DNA of an organism to the plasmid and create 'sticky ends'

Stage 2: Preparing a vector for the transferred gene

-Restriction enzymes are also put in the same container as the plasmids

-The same restriction enzymes that had cut the targeted gene from the DNA strand is used to create a cut in the plasmid, creating 'sticky ends' complementary to the DNA fragments from an organism

Stage 3: Recombinant DNA

-DNA ligase catalyzes the reaction and reconnects the cleaved plasmids with the DNA strands by creating covalent bonds

Stage 4: Isolation of transformed cells

-The recombinant DNA is then inserted into the host cell

-The cells that are transformed to contain the recombinant DNA are separated from untransformed cells