BUGSS:Build-a-BUG:2-3

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Build-a-BUG: Series 2, Session 3

Saturday, March 9, 2013
Noon to 4:00 PM

This is the third Build-A-BUG workshop in a series of five on yeast. You will learn more about molecular cloning and continue to hone your basic lab techniques, while continuing either your own or somebody else's mating type detector project.

Background reading

• Review Session 1 and Session 2
• Polymerase Chain Reaction (PCR) at NCBI

Polymerase chain reaction (PCR) protocol

• Mixture
  • PCR master mix - 45.0 μL
    • Nuclease-free water -35.6 μL
    • 10X PCR buffer - 5.0 μL
    • Forward primer (VF2) - 1.6 μL
    • Reverse primer (VR) - 1.3 μL
    • Deoxynucleotide Triphosphate (dNTP) Mix - 1.0 μL
    • Taq DNA polymerase - 0.5 μL
  • Template DNA (3002) - 5.0 μL
• Thermal cycler program - 30 cycles
  • Melt at 95°C for ? sec
  • Anneal at ?°C for ? sec
  • Extend at 72°C for ? sec

Materials

• Taq DNA polymerase
• Deoxynucleotide triphosphate (dNTP) mix