PCR from yeast colonies

[Michael Lichten]

Anonymous wrote wanting to know how to do PCR from yeast colonies.  We have 
routinely done PCR from single colonies; we use a 100 microliter PCR 
reaction volume and use an entire (2mm diameter) colony.  Just pop it off 
the plate with a flat toothpick, resuspend in the reaction mix, and cycle 
away.  No need to preboil or any such stuff.  What is important for 
reproducible success is to use FRESHLY GROWN colonies.  Even sticking the 
plate in the fridge overnight has given us problems.  If the colonies are 
more than 2 days old, you will get failures. 

Michael Lichten
lichten at helix.nih.gov