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Psilocyber's Syringe Tek v1.0
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 and sterile glovebox. There are many simple methods of
glovebox construction; most are available on the web at the popular
mycological culture websites. If you do not wish to construct a
glovebox, or do not have one, the following steps have been
performed with success by using the "oven tek." This simply involves
using your oven as a way of reducing the amount of contamination
present. To do this, turn your oven on its "warm" setting. Pull down
the door. You may then use the open door as a working surface or
pull out one of the oven racks halfway and work on it. The theory
(unproven) is that rising heat from the oven causes airborne
contaminates to rise and therefore prevents them from settling on
your working surface and therefore reducing the number of
contaminates present.
Materials needed: Empty
sterile syringes Two quart (or larger) saucepan One bottle of
91% isopropyl alcohol Several paper towels A lighter or
alcohol flame A shot glass Sterile spore print
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 cleans 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 syringe in a cool draft-free
place, preferable 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 door working surface: The shot glass, your
cooled syringes, the bottle of alcohol, a paper towel, your print
(still in zip-lock baggie) and the lighter or alcohol flame.
3. Wash hands with antibacterial soap before proceeding
further. 4. Fold the paper towel up into ¼ sections and
soak a corner of it with the alcohol. 5. With the alcohol
soaked towel wipe the interior of the shot glass, essentially
sterilizing 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. Then take your alcohol soaked paper
towel and wipe the needle with it to further aid in the
sterilization. At this point try to avoid letting the needle touch
any other surface unless otherwise instructed to do so.
7. NOTE: it is important at
this point to work as quickly 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 the needle of the syringe, 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 should notice that the water inside has turned a darker
shade of color and you may see small clusters of spores floating in
the solution. This is good, you have completed the process.
12. Sterilize the needle again with the alcohol soaked
paper towel, 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 in slow or no germination.
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