STUDY GUIDE FOR INCOURSE EXAM #1 DNA EXTRACTION AND AGAROSE

GEL ELECTROPHORESIS

1. Explain all the steps required for DNA extraction with justification for the use of
each reagent and each condition e.g. buffer use, detergent use, centrifugation steps
etc.

2. Explain the use of cold temperatures and buffers in the extraction of DNA.

3. Explain how DNA can be quantified using UV spectrophotometry.

4. Explain what methods other than UV spectrophotometry can be used to quantify

DNA.

5. Explain why purity determination is needed.

6. Explain how purity is determined.

7. What contaminants can be included in your DNA prep?

8. Explain the principles of agarose gel electrophoresis.

9. Explain the use and dangers of ethidium bromide.

10. Why must DNA be visualized?

11. Why does the 280nm wavelength used to measure.

12. What other molecules can be detected at the 260nm wavelength?

13. What factors affect the purity ratio?

14. State the equipment and reagents required to run agarose gel electrophoresis.