1. Last class Class policies, etc. PCR VNTRs

2. Visualizing DNA DNA is not colored; Can’t see in solution. Run mixture through an __________ solution is like JellO. Liquid when hot, solid(ish) when cold - Separate pieces based on

3. Separating DNA Agarose gel is porous ____________ in gel allow DNA to pass through ________ DNA passes easily, ____ DNA less easily

4. Agarose gel http://www.youtube.com/watch?v=2UQIoYhOowM

5. Loading dye Why do you need a “buffer?” Why do you add glycerol? Why do you need the “dye”?

6. Seeing the DNA EtBr + UV light = Gel/buffer contains ETHIDIUM BROMIDE (EtBr) EtBr in DNA + UV light = Regions in gel with DNA will show a BRIGHT “band” http://wikidoc.org/index.php/Agarose_gel_electrophoresis

7. Estimating size of DNA See DNA. But size? 200 bp 300 bp 400 bp 500 bp 600 bp 1000 bp 2000 bp http://wikidoc.org/index.php/Agarose_gel_electrophoresis

8. Changing gel parameters Increase voltage Increase percentage of gel Why ? Watch out! Increase time of run

9. LAB 2 Gel will be poured for you Load and run gel Check results Lab 3 onwards = pour your own gel

10. How do we study life? I think “X” BECAUSE “Y” Design an experiment to PROVE “X” Experiment:Predict outcomes if “X” is TRUE Predict outcomes if “X” is FALSE Do experiment, observe results Results; Therefore “X” is TRUE/FALSE Unexpected results/observations = Discovery!

11. Restriction endonucleases RE is an enzyme! - RE cutting is random!

12. Designing experiments Tube 10X Buffer (µL) RE (µL) Water (µL) 1N HCl (µL) 1 1.0 -8.00.0 2 1.0 7.00.0 3 1.0 -7.01.0 4 6.01.0 5 -6.02.0 61.0 5.02.0

13. Designing experiments Form a hypothesis Design an experiment Predictions?

14. LAB 2 Design experiment to test ONE variable - Time - Buffer - Enzyme concentration Discuss with partner, other groups & TA Generate experimental design Include controls!