pcr from colonies

[Georgiana May]

We use Lee's method (BioTechniques 15, Sept.1993).  We start with 3 small squares of mycelium, each about 1-1.5 mm sq. and trim away agar underneath.  These go into 0-ring Eppendorf tubes with 50 uL extraction buffer.  We follow the method and then use 2
uL/25 uL reaction.  It comes out about right for Coprinus cinereus, sometimes a 1:10 dilution works better.  The mycelium is freshly grown and the squares are removed about 1 cm back from the colony edge.

Georgiana
May