(Hayakawa & Nonomura, 1989) used the following procedure to re-activate dormant spores of actinomycetes:
- prepare either Yeast extract in PBS (6% w/v) or SDS in PBS (sodium dodecyl sulfate, (0.05% w/v), buffered to pH 7 by phosphate buffer.
- prepare inoculum in PBS buffer
- add 0.5 ml of inoculum in PBS buffer to 4.5 ml of enriched buffers
- keep on 40C for 20 min
- spread on plates (if making WAY, calculate how much yeast to add).
My modification – dissolved yeast extract and SDS directly in PBS, dilluted dry culture pieces in it. – DOESN’T WORK. Neither heat shock, nor nutrient shock, nor bleach shock work.
Sandye Adams: If re-plating from an old culture, simple streaking may not work. Take a chunk of the colony and put it in between two layers of agar. Or, do liquid culture.