Tom Schulz asks about generating =FBDNA from =FBsea star larvae for PCR. =
=FBWe=20
have successfully amplified from single urchin larvae, and have learned a=
=20
few tricks. Larvae can be placed directly into the PCR cocktail in a=20
microliter of sea water. A hot start helps prevent DNA degradation.=20
Better yet, we do a micro PK/RNase digestion by adding a larva in 1=20
microliter to 2 microliters of 100ug/ml Proteinase K, 10 ug/ml RNase.=20
This is incubated at 65 deg in the PCR machine for 1 minute, brought to=20
94 deg., and then the PCR cocktail added.=20
Good luck,
Steve Palumbi
