Yeast colony PCR – Department of Biology - University of Copenhagen

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Transcription, chromatin and DNA repair > Resources > Protocols > Colony PCR

Yeast colony PCR (up to 2 kbp).

  1. Using the tip of a toothpick, transfer a small amount of cells to the bottom of a PCR tube.
  2. Microwave for 1 min at max power together with a beaker of water (to absorb excess energy).
  3. Place on ice.
  4. Prepare reaction mix:   

     H2O   12.75 µl
     10xBuffer   2 µl
     20 mM MgSO4   1 µl
     2 mM dNTPs   2 µl
     Primers (20 µM)   2 µl (1 µl each)
     Taq (1 unit)   0.25 µl

  5. Mix on ice and add to PCR tube.
  6. PCR:

     96°C  2 min
     30 cycles of:  
     94°C  1 min
     52°C  30 sec
     72°C  2 min
     4°C  forever