Gel Interpretation & DNA Clean-up

1. Gel Interpretation&DNA Clean-up Tucson High School Biotechnology Course Spring 2010

2. What have we done so far? negative control psbA virus DNA positive control negative control DNA pol virus DNA positive control negative control g23 virus DNA positive control

3. What samples should we see a band for? negative control NO psbA MAYBE virus DNA positive control YES negative control NO DNA pol virus DNA MAYBE positive control YES negative control NO g23 virus DNA MAYBE positive control YES

4. Interpreting your gel psbA DNApol g23 Negative control Positive control Virus Negative control Positive control Virus Negative control Positive control Virus

5. What should you write about your gel?

6. Hypothesis Myovirus Podovirus X X psbA X DNA pol X g23

7. Next… Sequencing Need to Clean Up DNA PCR reaction taq polymerase dNTPs primers MgCl2 buffer DNA clean up DNA

8. MinElutepcr Clean-Up pcr products Buffer PE Buffer EB KEEP!! MinElute Column Buffer PB MinElute Column MinElute Column

9. Which pcr products will you clean up? psbA DNApol g23 Negative control Positive control Virus Negative control Positive control Virus Negative control Positive control Virus