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PCR Puzzle

PCR Puzzle. Class instructions. Start of lesson. Have the following at front of the class: Template. Start of lesson continued. Primers Bases. Correct base pairing is critical!. Green (Guanine) pairs with yellow (Cytosine). Blue (Adenine) pairs with orange (Thymine).

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PCR Puzzle

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  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?

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