1. PCR Puzzle Class instructions

2. Start of lesson Have the following at front of the class: Template

3. Start of lesson continued Primers Bases

4. Correct base pairing is critical! Green (Guanine) pairs with yellow (Cytosine) Blue (Adenine) pairs with orange (Thymine)

5. The devil is in the detail! The 5’ prime and 3’ prime ends of the bases must be round the right way!

6. Cycle 1 - denature Spilt class in 4 pairs or groups of students Each group will copy one STRAND from the template Thus, it is important to think of a template as two strands rather than one double stranded piece of DNA

7. Cycle 1 - denature Two groups will do the first cycle Give Group 1 one template strand Give Group 2 the other template strand

8. Students comes to front to get primer One student from each group comes to front of class to get primer

9. Students comes to front to get primer If they have double orange in their template, pick one double blue primer The other group takes one green-blue primer

10. Cycle 1 - Anneal primer Each group adds primer to the template Make sure 5’ to 3’ prime is correct!

11. Cycle 1 - extend Students take bases required to finish strand And add the bases to complete the copy

12. Cycle 1 - observations When finished filling in blanks, each group brings product to front of class and put on desk or floor

13. Cycle 1 - observations Make observations: – How many copies do you get? – How many are just the target sequence (ie begin and end with primers)?

14. Cycle 2 - denature Groups 1 to 4 will do Cycle 2 Give each group a strand:

15. Cycle 2 – Anneal primer Each group adds appropriate primer to the template strand Make sure 5’ to 3’ prime correct way round!

16. Cycle 2 - extend Students take it in turns to come to front to take matching base to add to template

17. Cycle 2 - observations When finished filling in blanks, each group brings product to front of class and put on desk or floor

18. Cycle 2 - observations Make observations: – How many copies do you get? – How many are just the target sequence (ie begin and end with primers)?

19. Cycle 3 - denature Groups 1 to 4 will do Cycle 3 Give each group two strands:

20. Cycle 3 – Anneal primer Each group adds appropriate primer to the template strand Make sure 5’ to 3’ prime correct way round!

21. Cycle 3 - extend Students take it in turns to come to front to take matching base to add to template

22. Cycle 3 - observations When finished filling in blanks, each group brings product to front of class and put on desk or floor

23. Cycle 3 - observations How many copies do you get? How many are just the target sequence (ie begin and end with primers)? What if anything, has happened to the template? How many bases with red, black and white do you now have?