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Registered: 11/06/04
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Dolly Lamb...sheep
    #3889682 - 03/08/05 11:15 PM (16 years, 2 months ago)

I want to clone soon..Was wondering if there was a different way to sterilyze the cardboard besides using the microwave?

"Take a canning jar and fill ? full with water. Screw a plastic lid prepared with a poly-fill wad on the top to filter incoming air drawn in while cooling. Place the trimmed cardboard wedge in the microwave and cook at boiling for 10-15 minutes. I achieve this by microwaving at full power until it boils, then turn the heat setting down. I find a low enough setting to allow cycles of boiling, then cooling without boiling over. You may need to experiment to find the ideal. This cooking is done to not only hydrate and sterilize the cardboard, but to cook the glue out as well. Upon cooling the jar is capped with an alcohol soaked coffee filter for transport back to the lab. Pressure cooking is not necessary with cardboard in my experience."



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Re: Dolly Lamb...sheep [Re: IGnosticAbhorI]
    #3889834 - 03/08/05 11:57 PM (16 years, 2 months ago)

Dr. Rush Wayne (the Peroxide Tek guy http://members.aol.com/PeroxyMan/ ) states you can sterilize with 1% peroxide (1 part 3% peroxide solution + 2 parts water). I've never tried it, but I know that would only be for mycelium, not spores. Here are his instructions:

How to prepare the plates

Here are the detailed steps for making cardboard plates. Note that you can also use small jars in place of Petri dishes.

1) Measure about 100 mls. of tap water into a small jar.
2) For nutrients, , measure another 100 mls. of tap water into a second jar and add one drop of ordinary soy sauce to the water, and a quarter teaspoon (1.25 mls.) of molasses or light malt powder.
3) Find some gray cardboard, the thicker the better, preferably gray on both sides. Trace a Petri plate onto the cardboard with a pencil and cut out several disks to fit into your plates.
4) Weigh one of your disks and record the weight. Multiply this weight by a factor of 1.3 as a rough guide (you may need to experiment with the amounts here), and add the resulting weight of tap water or nutrient solution to each disk in its Petri plate. (Remember, 1 ounce of water equals 28.35 grams; one gram equals one milliliter.) Example: Suppose my disks weighed 0.17 ounces each. Multiplying 0.17 by 1.3, I get 0.22 ounces. There are 28.35 grams in an ounce, so 0.22 ounces x 28.35 equals 6.3 grams. That means I'll add 6.3 milliliters of solution to each disk.
5) Close up the disks in the plates, and let the water or nutrient solution soak in.
6) Pressure-sterilize the jar of plain water, and the Petri plates with moistened newsprint disks inside, for 10 minutes at 15 psi (allowing the cooker to equilibrate steam for 10 minutes before putting on the pressure regulator).
7) Cool the cooker, and remove the plates and jar of plain water.
8) When the water has cooled, add 3.3 mls. 3% peroxide to the jar, using a pasteurized pipette, to give you a final concentration of about 0.1% peroxide in sterile water.
9) Add about one third of the initial weight of the cardboard as 0.1% peroxide to each disk. Let the solution soak completely into the disks. They are now ready to use.

You can store and incubate these plates inside plastic food storage bags as I suggest in Volume I for agar plates. But you will probably find that your cardboard disks dry out too quickly. You can keep them moist longer by storing them in a closed container that has some peroxide solution in the bottom. For instance, find a plastic yogurt container or a jar with a mouth wide enough to let Petri dishes pass. Then create a platform to hold your Petri dishes off the bottom of the container, perhaps by putting a smaller jar inside the larger container. Put the Petris on top of the platform. Then add a small quantity of peroxide solution to the container at the same concentration you use for your plates (roughly 0.018%). Finally, cover the container with a layer of plastic wrap and fix it in place with a rubber band around the mouth of the container (this kind of closure will allow adequate oxygen diffusion). Be careful to set it up so that you cannot knock your Petri dishes off the platform into the water.

- Mages

Ignorance killed the cat; curiosity was framed.
- C.J. Cherryh

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