Yeast colony PCR (up to 2 kbp).
- Using the tip of a toothpick, transfer a small amount of cells to the bottom of a PCR tube.
- Microwave for 1 min at max power together with a beaker of water (to absorb excess energy).
- Place on ice.
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
- Mix on ice and add to PCR tube.
96°C 2 min 30 cycles of: 94°C 1 min 52°C 30 sec 72°C 2 min 4°C forever