Yeast colony PCR

1. Resuspend a swath of yeast (about the size of a glass bead) in 12µl lysis buffer.
2. Incubate at 37°C for 5 min, then 95°C for 5 min.
3. Dilute the DNA 5-10 times with water, and use 3µl for 50µl PCR.
   

lysis buffer:

• 0.1 M NaPO₄ buffer of pH 7.4
• 2.5 mg/ml Zymolase 20T (21100 U/g, Seikagaku Corporation)

Note: It is important to use very fresh yeast. At least use yeast grown O/N on YEPD plates. I grow yeast first on -Leu -Trp for 1 or 2 days, then replica plate to YEPD and use them the next day.