Nazarbayev University

• BIO 110L

Laboratory #6 Plasmid Mapping Aim: The aim of the study is to understand the principles behind plasmid mapping with the usage of restriction enzymes Sample Plasmid Maps Plasmid S2 Modern Biology I laboratory Plasmid Map Questions: 1. From the map of plasmid S2 list all the restriction enzymes that would cut this plasmid. PvuII, Scal, EcoRI, BamHI, Pstl, and HindIII 2. Which plasmid, S2 or S5, is the biggest and what is its size? S5 is the biggest, because it has 9481 bp, while S2 has 5869 bp. 3. Using plasmid S2 as an example, find the restriction sites for the enzyme PvuII. How many sites are there? What is their location? If PvuII was used to cut (digest) this plasmid, how many fragments would it make? 3 sites for enzyme PvuII. Positions – 55, 1993, and 3410 bp. 3 cuts, that’s why 3 fragments will be produced. 4. Next determine the size of the fragments created when plasmid S2 is cut by PvuII. DNA fragment size is calculated by subtracting the site locations from each other. (Note: if a fragment contains the 0 point of the plasmid, it is not just a simple subtraction!). How big are the fragments from plasmid S2 that is cut with PvuII? The fragment sizes should add