Biology
Biology Lab: AP Investigation #9 — Restriction Enzyme Analysis of Bacterial DNA
Lab Exercise
Determining Bacterial DNA Sequences
Concept: Bacterial DNA can be manipulated by insertion of genetic material. Bacteria will express inserted genetic instructions in ways that can be detected.Goal: To become familiar with and practice methods of genetic manipulation.
Read through the 2012 AP Biology Lab Manual, Investigation #9: Restriction Enzyme Analysis of DNA. This lab presents signification problems for home students in acquiring DNA samples and restriction enzymes, but is worth looking at to see the technology involved.
If you are unable to obtain all the equipment and materials required by Investigation #9, you should still be familiar with the AP Lab as designed, please review that investigation in your lab book.
Another good review of enzyme restriction to create bacterial vectors is given in the Addgene Restriction Enzyme article. Addgene is a non-profit organization allowing biologists to share plasmids they create in individual research projects.
If you still want to perform this lab in some way, you can further your DNA extraction skills by performing the DNA extraction lab below. You will need the gel electrophoresis equipment and dye materials you made using Illustrated Guide to Home Biology Experiments Labs III-6 and III-7 to sort the DNA you extract. Then complete the simulation lab using DNA restriction enzyme techniques to analyze DNA evidence and identify which suspect is guilty. Follow the instructions, and note your materials and procedures as though you were actually performing the lab. Record your data, give examples of your data analysis, and write up your investigation as a formal lab report.
- Extracting DNA from Strawberries (or other sources). If you decide to use blood from any source, be sure to use appropriate precautions, including wearing examination (latex) gloves to handle all samples, to avoid both contamination of your samples and prevent infections for the lab investigators!
- The Genetics Science Learning Center has instructions to help you Build a Gel Electrophoresis Chamber.
- There is a gel electrophresis simulation at the Lab XChange which can help you learn how to use the chamber you build.
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