pick entire colony and resuspend
in 100 ulsterile water.
use 2-5 ul for standard
25 ul PCR reaction with Taq pol, run 10' soak at 94 C before cycling
use the rest of the suspension
to start cultures of clones that scored positive in the PCR reaction
PCR products produced this
way can be sequenced or cloned without any further cleanup.
PCR products can be used
as template for in vitro transcription without cleanup if you ad
RNasin (the most abundant E. coli RNases are effectively inhibited by RNasin)
boil yeast colony in PCR buffer for 10', then run normal 25 ul PCR reaction with Taq pol.