Easy syringe method

Nan's Nook : Archives : Spores : Syringes : Easy syringe method
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By Vitti (Admin) on Thursday, August 23, 2001 - 12:06 pm: The Nook

Bring some water to boil on the stove and let it boil about 10 minutes.
Suck the boiling water up to a fresh syringe.
If the syringe was used before, sterilize the allready filled syringe in a PC for 15 minutes.
Put syringe aside until cool.
Following tasks should be idealy prepared in front of a flow hood, but a reasonably cleaned bathroom will do:
Prepare the print, a shot glas and the alcohol lamp.
Rinse the clean shotglass with isopropanol or rubbing alcohol and let it evaporate a few seconds.
Bring the needle of the syringe to red glow above the alcohol lamp.
Squirt 1/2 ml of water out of the syringe to cool the needle and create some airspace in the syringe so later you can shake the spores.
Shake of the eventual drop on top of the needle off so the print won´t become wet.
Scrape some spores with the tip of the needle into the shotglas.
Add a small amount of water from the syringe, stir and suck it back into syringe.
Give the syringe a vigorous shake, let the spores rehydrate for a few hours and inoculate

By Admin (Admin) on Thursday, August 23, 2001 - 12:06 pm: The Nook

This is how my monkey does his syringes every time...sterilizes shot glass in microwave(about 5min-hi) , he does this by putting his shot glass in a cup and filling the cup with water- then boiling it down so the shot glass can be removed with fingers once it's cooled ... he sometimes puts a bowl on the bottom to catch the splatter inside the microwave {Bowl-pyrex measuring cup[filled to 1cup capacity]-shot glass}boils some water, let's that cool... now the water that's left in the measuring cup can be used as syringe water- or if your syringes have been cooling because you just cleaned them (and their full of water) then you have plenty of water to make a syringe...
now ... what monkey does is pretty risky in some eyes in that he has this little home made innoculating loop... basicly what it is is a cheapo knife with the tip bent into a loop.. he then proceeds to scrape as many spores as he wants to into the shot glass, squirts out the contents (h2o)of the cooled syringe, sucks up and squirts out again to clean any of the spores that may have stuck to the sides.. sometimes he repeats this a couple of times if there aren't that many spores to be frivilous with.... i hope this helps out ... i think it would be easily applied to a jar or what ever glass container you have- that is , if you have a glass containter that is larger than it so you can sterilize it in the microwave

By Nan (Nanook) on Monday, October 22, 2001 - 12:37 am: The Nook

How to Make a Spore Syringe


Sterility is key. If your spore print is contaminated or you introduce contamination into the spore syringe, you will have difficulty later in the process. Ideally, there should be no fans blowing or drafts of air. You should clean the area where you will be working carefully and make sure that everything is tidy.
Materials needed:

spore print
10 cc. syringe with long needle Where to find
shot glass
regular glass
coffee mug
cigarette lighter
X-Acto knife or sharp steak knife
microwave oven
Several things need to be accomplished. First, we need to sterilize a shot glass to easily mix the spore solution and we need to sterilize a syringe to hold the solution. We also need some sterile water in which to suspend the spores. The following procedure will accomplish all of this.

Fill a coffee mug with water and place a shot glass inside the coffee mug. Make sure the shot glass is completely submerged. Place the coffee mug in the microwave oven and get the water to a full boil for 10 minutes. It does not need to be a violent boil. Adjust the heat level of the microwave oven to keep too much water from being lost if necessary.

Remove the shot glass and empty the excess water out of the shot glass. Place another glass over the shot glass. This will keep air born contaminates from settling in the shot glass while you wait for the shot glass and water in the spore syringe to cool.

Fill the syringe with hot water from the mug. Eject the hot water and repeat several times. This will insure the inside of the syringe and the needle are clean and sterile. This is especially important if you are using a syringe from a previous crop. When the needle is inserted into the substrate, it is possible to get nutrients up inside the needle and for contamination to grow. The last time you fill the syringe with hot water, do not purge it. Let it sit in the syringe until it is cool. This is useful for two reasons. First, the continued heat from the water can still work to eliminate any remaining contaminates. Secondly, once the water is cool it can be used as the sterile water needed to fill the syringe. Make certain that nothing touches the needle of the syringe.

The Psilocybe mushroom spores will be killed if they come in contact with anything too hot. You need to wait until the shot glass and spore syringe are at room temperature. When it is safe to proceed, use the cigarette lighter to flame sterilize the X-Acto knife and the needle of the syringe. Let the blade of the knife cool, but make sure it does not touch anything. When it is cool, carefully open the spore print and scrape a fleck of spores into the shot glass. A fleck 1/4 inch by 1/4 inch is more than sufficient for a 10 cc. spore syringe. Use the needle of the syringe to stir the spores into a few drops of water. Usually, there will be a few drops left over in the shot glass from when you emptied it. Otherwise, you can get the drops from the syringe. Stir the fleck of spores until they are well broken up and 'dissolved' into the water. Purge the water out of the syringe into the shot glass. Pull the water back into the syringe, being sure to suck the spores in also. Do this once or twice more to make sure the spores are well mixed in the spore syringe. Often, it takes several tries to get the spores fully broken apart and well mixed.

If the spores in the print have been dried and are not fresh, it is best to wait six hours to use the spore syringe. The spores need to rehydrate. If your in a hurry, the spores can still rehydrate in the culture jars.


How to Make a Spore Print


Once you have a mature mushroom, you are in a position to make a spore print and use it to continue cultivation of mushrooms. The cap should be harvested when the mushroom cap has become flat or is starting to invert.
Sterility is key. Adaptation-25 Be careful not to do anything that will compromise the sterility of your spore print. The typical procedure is to cut the stalk off of a mature mushroom very close to the cap. A sterilized knife or razor blade is used to do this. The cap is then laid on a sterile piece of tissue paper or card stock and a small glass set over it. The glass is needed for two reasons. First, it keeps the spore print insulated from airborne contaminates. Secondly, it helps keep the humidity high so the mushroom cap can continue to live and drop its spores. One note of caution. Some humidity usually needs to be allowed to escape. You want the environment inside the glass to be slightly less humid than the environment in which the mushroom was grown. If you have problems getting a cap to drop its spores, try using a piece of paper for the print that fits entirely inside the glass and spreading out a wash cloth flat on the table. Let the edge of the glass seal to the wash cloth instead of the paper. This will usually allow enough humidity to escape to cause the cap to drop spores.

If everything goes well, after a day or two the cap will drop its spores. Adaptation-26 There will be a purple-brown dust underneath the cap. These are the spores.

Eye glass lens paper is good source of sterile tissue paper. A box of waxy tissue paper that deli's use to pick up donuts and rolls is another excellent source of sterile tissue paper. Card stock (such as a recipe card) is a bit easier to use later when you want to prepare a spore syringe, but you have to expend the extra effort up front to sterilize it. To use card stock, place in a 425 degree F. oven for 15 minutes. Let it cool with a glass over it before you place the cap on it.
Once you have a spore print on the paper, remove the glass and cap. Fold the spore print in half and seal the edges so air can not get in. A piece of scotch tape on each side will do nicely. The spores will stay viable for 18 months if they are kept in a cool, dry and dark spot. If you place a small amount of desiccant in the bottom of a film container and place a cotton ball on top of the desiccant, you have an ideal container to keep the spore print. The cotton ball will keep the desiccant from touching the spore print. Seal the spore print in the canister and place the canister in your refrigerator until you are ready to use it.

Note that if you want to be self sufficient, it is a good idea to have multiple spore prints and store them separately. You just never know when you are going to be surprised with a massive contamination problem or thermal death. The safest thing to do is have a couple viable spore prints so it is easier to recover from disaster. A spore print is typically viable for about a year if it is stored in a cool, dark location. As a print ages, germination gets slower and this process becomes more prone to contamination.


Combination of Spore Print and Preparation of Inoculant


This excellent idea is from: AN165023

If it is your intent to use a mushroom to immediately generate inoculant for a successive crop, you can eliminate some of the above steps and reduce the risk of contamination.

Poke a small nail hole in the cap of a wide mouth jar. Cover the nail hole with a piece of electrical tape. A 1/2 pint canning jar similar to that used for the cultures is fine. Sterilize both the jar and the lid.

Place the harvested cap in the sterilized, wide mouth jar. Seal the top with the lid and wait until a spore print has been generated on the bottom of the glass. Open the jar and remove the mushroom cap. Add 3/4 cup of sterilized water. Seal the top of the jar with the lid and shake the jar. The spores need to be mixed well into the water. This procedure elements the need to transfer spores from a print to some container to make the solution. The spore print is generated inside the container and the only extra step is adding water. It also has the benefit of making a large amount of solution that is easy to use.

Now, any time you need inoculant, you can shake the jar and peel the tape back to expose the nail hole in the lid. Simply insert the needle of the syringe into the inoculant and pull some into the syringe. The syringe needs to be sterile or you risk contaminating the entire jar of solution. The solution will keep longer if you refrigerate it. You still should keep some spore prints on paper because it is possible to contaminate the entire jar of inoculant if you make a mistake.

By Cbee (Cbee) on Tuesday, December 11, 2001 - 03:40 am: The Nook

Im about to try to make some syringes from my PRs. Now I understand all the stuff about a glovebox but i havent had time to make one. Can i still make a sterile syringe without one? I am planning on doing the trick nan posted in the archives about puttin a rack that rests from teh top of the jar and then the spores drop right into the water. I am planning to do this:

take a half pint short jar and put in the metal wire to set cap on and fill with some water. Put a plastic lid on and through into the steamer. Next without opening the jar take it to the room my terrarium is in. The put the closed jar (cooled) near the cake and snip the cap with sterilized scissors and hold it with a sterilized pin. The open the jar lid as little as possible and slide the cap on the wires. The screw lid back on and wait 2 days... think this will be sterile enough to get viable syringes? The lid will have holes in it, but taped up during this whole process. Then when i want to make my dozen + syringes Ill open one hole and draw one all the solution i need.. I think this will work without a glovebox, or should i wait for a later flush when I have a glovebox. The monster I wanna get the spores from will be opening tomorrow so please any quick suggestions would be great =) thanks alot everyone

By jack cracker (Cptcracker) on Tuesday, December 11, 2001 - 04:35 am: The Nook

if i were going to go through all the change to make syringes, i would be sure to do it in at least a Clean Room, if not Sterile...
a simple glove box is in the archives, made with a Cardboard box!
using the Oven as a Flow Hood is pretty effective too, also in the Archives...
there is also a little reading on clean room techniques there...
i guess if i'm going to point i should learn to link, d-oh.

By Nan (Nanook) on Tuesday, December 11, 2001 - 05:35 am: The Nook

Follow 9er Tek or PF Tek and you will be fine.
By Cbee (Cbee) on Thursday, December 13, 2001 - 12:36 am: The Nook

ok i made a jar like that with wire and shit, and spores have dropped over night, and im goina leave it till tomorrow, how do i take the cap and wire out without introducing contams? and do i just boil the syringes for an hour to sterilize? i think what i may do is suck up the water with the cap and wire still in, then i wont have to open the lid at all, thus reducing contams

By quote: (Quote) on Thursday, December 13, 2001 - 12:50 am: The Nook

work inside a glovebox.
failing that, try the oven door method when you remove the cap. and, of course, while you prepare syringes.
20 minutes boiling is adequate for syringes.

By Cbee (Cbee) on Thursday, December 13, 2001 - 08:11 pm: The Nook

cool, thanks quote, i am goina try to make a glove box today

By Liberty_Caps (Liberty_Caps) on Saturday, January 05, 2002 - 11:39 pm: The Nook

if im going to make some more PRINTS so i was curious.......
is it better to scrape all the spores in the water so thier like dust?????? or will that damage them??????? can you damage spores by scrapeing them????? because my syringes are black but have no spore chunks in them...
it's like dye in water???
jut wondering.....
is it better to have chunks?

* 1) Scraping is fine, you do not need to scrape the entire print usually 2) No damage 3) Like dark powder in water
4) Powder - Nan

By quote: (Quote) on Sunday, January 06, 2002 - 01:28 am: The Nook

no, you don't want chunks.
powder is best.