The goal of Plasmid Fingerprinting is to cut your plasmid into specific sized pieces and analyze the resulting fragments by gel electrophoresis

The goal of Plasmid Fingerprinting is to cut your plasmid into specific sized pieces and analyze the resulting fragments by gel electrophoresis. The pattern of the fragments on the gel is determined by the number of cut sites for a specific enzyme and the location of those cuts. By selecting the appropriate enzyme(s), one can either linearize a plasmid to determine the size of the entire construct or cut it into many small fragments.

Treat each of the following digests as individual applications.

Consider a digestion of the pET28a plasmid with the following enzymes. For each digestion, (1) list the cut site sequence, and (2) estimate the size(s) of the linear DNA fragments that would result. Keep in mind that pET28a is 5369 bp.

1. a. EcoRV – Cuts at 1573
2. b. Acc1 – Cuts at 180 and 2994
3. c. BspH1 – Cuts at 521, 3944, and 4819
4. d. BsiE1 – Cuts at 169, 1908, 3140, 3564, and 4426