In this lab students will understand the action and specificity of restriction enzymes, while learning the components involved in the process of DNA electrophoresis and the role of each component as it applies to the entire process of electrophoresis. Students will perform the electrophoresis process on DNA samples treated with different restriction enzymes and construct a standard curve using a known DNA sample while determining the approximate size of the DNA fragments in unknown samples.
Materials provided for eight lab groups.
Meets AP Science Practices 3 and 6, and Big Idea 3.
Prepared agarose, 0.8%, 200ml TBE buffer, 5X, 500ml
DNA Stain, 20X, 60ml DNA Samples, 100ul each:
DNA Marker (Lambda DNA HindIII digest)
Sample #1 (Lambda DNA)
Sample #2 (Lambda DNA BstEII digest)
Sample #3 (Lambda DNA EcoRI digest)
Sample #4 (Lambda DNA BstEII digest)
Materials Needed but not Supplied:
Agarose electrophoresis chamber Microwave or hot water bath
Electrophoresis power supply Distilled or deionized water
Micropipettes w/micropipette tips, 10μl
Gel staining trays or similar (large enough container to accommodate flat agarose gels)