Liquid psilocybe spore syringe
STEP 1. Day 1. How to prepare liquid spore syringe without glove box
For now you've already prepared everything you need according to the lists of stuff for growing process.
The first step in cultivation process will be liquid spore syringe preparation for further substrate inoculation.
*Skip this step if you already have a ready to use liquid spore syringe and move on STEP 2. Day 1. Choose and prepare substrate for magic mushrooms
What is liquid spore syringe?
Liquid spore syringe — consists of a syringe filled with mushroom spores in sterile water for spores hydration (rehydration). It is one of the method for transporting spores to a sterile substrate (inoculation). Spore syringe is the fastest and easiest way for inoculation.
Spore syringe VS Liquid culture
Spore syringes are often mistaken for liquid culture. Liquid culture is the result of an isolated mycelium culture grown and expanded in a sugar or honey broth.
Liquid culture (LC) is preferred over a spore syringe to inoculate grain spawn due to the reduced chance of contamination. But spore syringe preparation can be a part of LC preparation. LC is more complicated process in comparison with spore syringe and needs more time.
Why we make a liquid spore syringe?
Let's turn to nature for the answer, because the whole process of mushrooms cultivation is a simulation of natural conditions.
When mushroom cap are opened, the mushroom begins to spread spores. Such fresh spores are ready to germinate, they contain enough moisture. But after a short time they become dehydrated and the cell walls of the spores curl toward the center. So that, they go into a dormant state for a period of drought and wait for the rainy season.
In order to reproduce the rainy season, thereby awakening and preparing spores for vegetation, we place them in sterile water in a syringe.
In addition, spore syringe is the easiest and most accessible method of substrate inoculation for newbie growers.
Other inoculation methods we'll discuss in the inoculation section.
Video "Liquid spore in zip bag without glovebox"
The following method of preparing the liquid spore syringe is the easiest and very simple. Moreover, there is no need to use a glove box or SAB. However, it is rather wasteful, because you need to sacrifice the whole print at once. At the same time it doesn't require perfect sterile conditions.
In this section we'll discuss how to make a larger volume of liquid spore syringe by diluting it. And how to store ready to use spore syringe for a long time.
In general, the process of preparing the liquid spore syringe is demonstrated in the video below:
Don't be lazy to read the detailed description of the process, because the video shows only the main principles!
List of required stuff for liquid spore syringe preparation
To prepare a liquid spore syringe on your own, you need:
1️⃣ Spore print in a zip bag. Ideally to have a sterile spore print. Running a little ahead I'd like to note: If you are not sure either spore print is sterile or not then antibiotic (gentamicin or any analog) should be added to the spore syringe before inoculation. If the spore print is sterile, then antibiotic is not needed. All details we'll be considered at the Inoculation step.
2️⃣ Sterile syringe. One syringe for one spore print.
The syringe volume is at least 5 ml. You can use syringe of medium and large volume of 10-20-30 ml, then you can dilute the liquid spore suspension and get more material for inoculation. I'll explain how to do this later.
3️⃣ Ampoule with sterile water for injection (5 ml or 10 ml). 1 ampoule per 1 spore print.
If you will be diluting the liquid spore suspension, you will need additional water for injection. Another 5-15-25 ml for 10-20-30 ml syringes, respectively.
4️⃣ Ampoule cutter or kitchen knife.
5️⃣ Ethyl alcohol 70% in a bottle or in a spray bottle.
It is better to use a 70% alcohol, but 96% is also okay. Some growers use antiseptics or chlorhexidine instead of alcohol. We don't recommend any substitution for alcohol. We use only alcohol (70%) without any additives. You can use 96% alcohol. Actually 70% is even more reliable, because it penetrates better through the membranes of the bacteria.
6️⃣ Cotton pads — 2-3 pcs.
7️⃣ Grower's outfit: gloves, face mask, cap, medical gown (or PE raincoat).
8️⃣ Table and chair.
Preliminary preparation before making liquid spore syringe
- Put on the necessary grower outfit: face mask, medical cap, gloves, medical gown (or PE raincoat).
- Wipe the table with alcohol (by using alcohol in bottle or spray bottle and cotton pads).
- Spray alcohol on your gloves. Wipe your hands well over the entire surface of the gloves.
- Spray alcohol on the sleeves of the medical gown (or raincoat) up to the elbow.
- Use alcohol to wipe down all stuff on the table: the ampoule with water for injections, zip bag with the spore print, the ampoule cutter (or kitchen knife), the syringe package and even the bottle with alcohol.
- Don't spare the alcohol!
Step-by-step liquid spore syringe preparation
During further manipulations don't hang over the table with the whole your body. Try to have only your palms and hands above the table.
1️⃣ Open the ampoule with sterile water for injections.
2️⃣ Open the syringe pack. Put the needle on. Fill the syringe with sterile water from the ampoule.
3️⃣ Don't open zip bag with spore print! Wipe zip bag with alcohol again.
4️⃣ Pierce zip bag closer to the valve. Put the needle inside. Squeeze all water from the syringe into the zip bag.
5️⃣ Don't pull the needle out of the zip bag. Just clamp the puncture point with one hand and rub the spore print through the zip bag with the other hand. So that, spores separate and mix with water.
6️⃣ Draw water with spores from zip bag back into the syringe.
7️⃣ Remove the needle from the zip bag. Wipe the syringe needle with alcohol (wipe by cotton pad soaked with alcohol or just spray it with alcohol). Put the protective cap over the syringe needle. Make sure opened syringe needle doesn't touch anything. Do not put the syringe on the table without the protective cap on the needle!
8️⃣ Shake the syringe to mix water and spores. Wrap the syringe in foil to ensure dark conditions.
9️⃣ Sign the strain.
🔟 Leave prepared liquid spore syringe to rehydrate for 24 hours at room temperature (+20+25°С or 68-77°F).
If the spore print has been stored for 2 years or more a longer spore rehydration will be necessary. Each year of storage increases the duration of rehydration by 1 day. 2 years of storage is 2 days of rehydration, 3 years is 3 days of rehydration, etc. Or you can just leave spores in water for a week. Then liquid spore syringe should be stored in the refrigerator at +3°..+5°C or 37-41°F during such long-term rehydration.
How long liquid spore syringe can be stored?
Up to 1 year liquid spores stay viable.
Liquid spore syringe should always be wrapped in foil and stored in the refrigerator at +3°..+5°C or 37-41°F.
Shake the syringe thoroughly before inoculation, because spores may stick together over time.
How to get a larger volume of liquid spore? How to dilute it?
You can dilute the prepared liquid spore, thus increasing the volume of raw material for inoculation by several times. This way, you can inoculate more jars of substrate. After all, each spore print contains millions of spores (about 3-5% are capable of continuing life).
To make the dilution as sterile as possible and to avoid contact of the liquid spore with other vessels, from the very beginning take a medium or large-volume syringe to prepare the liquid spore. For example, 10-20 ml or even 30 ml.
Make all manipulations for liquid spore syringe preparation in zip bag. Then simply add another 5-15-25 ml of sterile water to the available 5-10 ml of water with spores in the syringe.
You can dilute the liquid spore until the solution is slightly stained with spores.
It is better to dilute the suspension just before inoculation. Then you can save on the antibiotic and you can store liquid spore for a long time.
Necessary antibiotic (gentamicin) concentration is 0.5 ml per 5 ml of liquid spore for 20-30 minutes before inoculation. If you dilute immediately, you will have to increase the antibiotic consumption accordingly: 1 ml per 10 ml of the suspension, 2 ml per 20 ml of the suspension, and 3 ml per 30 ml of the suspension.
If you add antibiotic you need to use the whole liquid spore at once. Because antibiotic for a long time is harmful for spores.
If you have a sterile spore print, antibiotic is not necessary. In this case, it doesn't matter when to dilute the liquid spore. You can do it immediately, or you can do it 24 hours later just before inoculation.
We'll discuss all details about using antibiotic in liquid spore in the Inoculation section.
While the spores will rehydrating for the next 24 hours or even more don't waste your time and move on to the next step — STEP 2. Day 1. Choose and prepare substrate for magic mushrooms ▶️
Feel free to ask any questions and leave your comments ⬇️⬇️⬇️