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Tuesday, August 12, 2008

moss culture - keeping sterile

Subject: BRYONET: Re: regarding moss culture
Date: Mon, 26 Jul 2004 11:59:21 -0400 (EDT)
From: Janice M. Glime <jmglime@mtu.edu>
To: jakob raath <lungumabel@yahoo.co.uk>

BRYONET

Dear Jakob,
I would sugest you make two changes in your protocol. First, do not
open the plates in the greenhouse. It will have lots of fungal spores and
sufficient air movement to contaminate your cultures. If at all possible,
take them to a clean room and open them in a sterile area. If this is not
practical, clean an area of the greenhouse with disinfectant and put up a
barrier around your working area to reduce air movement while you work.
The best solution would be to make a small hood with a plexiglass front
and two holes for your hands. Make a cloth or other flexible collar for
the opening so it makes a seal around your hands when you work with your
plate inside. You can put a microscope inside if you need and make a
small hole for the oculars. For added protection, you can install a UV
bulb and white light bulb (fluorescent). Turn on the UV when you are not
working and the white light when you are. You may need to disinfect this
chamber before each use.
My second suggestion, which you might already be doing, is to use a
non-nutrient agar. Put only mineral nutrients in the agar so there is a
low nutrient and as little as possible carbon source for fungi and
bacteria. Mosses need much lower nutrient concentrations than other
organisms.
You might need to transfer your cultures every 2-3 weeks. Fill the
plates almost to the top to maintain moisture longer.
Seal the plates with tape or preferably parafin film that you replace
after you open it.

Janice
***********************************
Janice M. Glime, Professor
Department of Biological Sciences
Michigan Technological University
Houghton, MI 49931-1295
jmglime@mtu.edu
906-487-2546
FAX 906-487-3167
***********************************

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