NOTE: These instructions are most effective
when performed in the most sterile environment available. The preferred
method involves following the steps below while working in a clean glovebox. There are many simple methods of glovebox construction; most are
available on the web at the popular mycological culture Websites, or in the Shroomery's
glove box section.
Materials needed:
Empty sterile syringes
Two quart (or larger) saucepan
One bottle of 70% isopropyl alcohol
Several paper towels
A lighter or alcohol flame
An
inoculation loop or equivalent (you can make one with thin wire)
A shot glass
Procedure One: Making a sterile syringe
1. Fill your saucepan halfway with tap or distilled water (use distilled
water if your tap water contains higher levels of minerals and chemicals).
2. Boil the water in the saucepan on high for a minimum of ten minutes,
this should be adequate to sterilize and cleanse the water of all bacteria
and viruses.
3. Take your empty syringe and fill it with the boiling water. Allow
it to sit for two minutes with the hot water inside.
4. Purge the hot water from the syringe into a sink, not back into
the saucepan.
5. Repeat steps 3 and 4 two more times. Upon the second time leave
the hot water in the syringe.
6. Place the capped syringe in a cool draft-free place, preferably in a
clean zip-lock bag
7. Allow it to cool for several hours before proceeding to Procedure
Two.
Procedure Two: Transferring print spores into
syringe
1. First clean your work area. This may involve wiping down all work
surfaces with a diluted bleach solution and spraying the area liberally
with a disinfectant such as Lysol.
2. Place the following materials in your glovebox or on the oven
lid working surface: The shot glass, the bottle of
alcohol, a paper towel, and your print (still in zip-lock baggie). Keep your syringes, inoculation loop, and flame outside the box.
3. Wash hands with antibacterial soap or isopropyl alcohol before proceeding further.
If possible, wear latex/nylon gloves.
4. Fold the paper towel up into ? sections and soak a corner of it
with the alcohol.
5. With the alcohol soaked towel wipe the interior of the shot glass, sanitizing the surface you are about to use in the transfer.
Allow the shot glass to air dry, should only take a few seconds.
6. Remove the needle guard from your sterile syringe and flame sterilize
the needle. Also flame-sterilize your inoculation loop, if possible (i.e. it's made out of metal). Then move the syringe and loop inside the glovebox. At this point try to avoid
letting the sterile instruments touch any other surface.
7. NOTE: it is important at this point
to work as smoothly and efficiently as possible to help combat the chances of contaminating
molds and bacteria falling into your work area and thereby ruining your
syringe.
8. Remove the print from its storage baggie. Unfold it to expose
the spores. Lightly begin to scrape, using your inoculation loop, a
section of the print off into the shot glass. For a medium sized print it
is usually adequate to scrape off a section no larger than 1/5 of the total
print.
9. You will have a small noticeable collection of spores in the shot
glass. Now expunge no more than half of the water from the syringe into
the shot glass, lightly stirring the spores into the solution.
10. Suck the spore water solution into the syringe. You may need
to expunge some more water into the shot glass and re-suck to help in capturing
all the spores into the syringe.
11. Once you have the spore solution back into the syringe you may notice that the water inside has turned a darker color and/or you
may see small clusters of spores floating in the solution. This is good,
you have completed the process.
12. Sterilize the needle again,
replace the needle guard and place the syringe back into your clean zip-lock
bag.
13. Allow the syringe to sit for no less than 12 hours before using
it in jar inoculation. This is extremely important, as the spores must be
allowed to rehydrate before they can be introduced into the substrate material.
Failure to allow this may result is slow or no germination.
*** Updated by andymc 20/01/11 ***
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