You will need the following items: 1  Temporary Use Permit Application (completely filled out). If the applicant is not the owner of the subject property, provide a Letter of Authorization with original signature. 2  Burden of Proof.
This is known as enzyme digestion. On double stranded DNA the recognition sequence is on both strands, but runs in opposite directions. This allows the enzyme to cut both strands. Sometimes the cut is blunt, sometimes the cut is uneven with dangling nucleotides on one of the two strands.
Restriction Enzymes Cut DNA Molecules at Specific Sequences. To clone specific DNA fragments in a plasmid vector, as just described, or in other vectors discussed in later sections, the fragments must be produced and then inserted into the vector DNA.As noted in the introduction, restriction enzymes and DNA ligases are utilized to produce such recombinant DNA molecules.he phenomenon of transformation permits plasmid vectors to be introduced into and expressed by E. coli cells. In order to be useful in DNA cloning, however, a plasmid vector must contain a selectable gene, most commonly a drug-resistance gene encoding an enzyme that inactivates a specific antibiotic.