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 (for example Restriction Endonuclease) are used to cut across the two strands of DNA and leave loose ends to which the cDNA can be attached. Special linking DNA sequences are added to connect the plasmid DNA with the human insulin cDNA.