STEP 1. Let’s make liquid spore syringe from a spore print. Lifehack for noobs: sterile spore syringe in zipbag without glovebox, SAB or laminar flow hood

Spore syringe preparation
How to make spore syringe

How to make Mushroom Spore Syringe without glovebox or SAB

STEP 1. Day 1.

For now you've already prepared everything you need according to the lists of stuff for growing process.

The first step in cultivation process is mushroom 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 anyway spore syringe preparation is 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 skip detailed guide below, because 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.

Magic mushroom liquid spore syringe preparation
Psilocybe cubensis liquid spore syringe preparation

2️⃣ Sterile syringe. One syringe for one spore print.

Disposable sterile syringe with needle
Disposable sterile syringe with needle

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.

Shroomok's Choice on Amazon:

5 ml/cc Disposable Sterile - 22Ga Needle - 10/20 Pack - $8.90

10 ml/cc Sterile Syringe - 18Ga Needle - 10 Pack - $10.90

20ml/cc Laboratory Syringe - 20ml with 21Ga Needle - 10 Pack - $9.99

3️⃣ Sterile injection water in vial (minimum 5 ml).

Sterile injection water in vial and sterile syringe for liquid spore
Sterile injection water in vial and sterile syringe for liquid spore

If you will be diluting the liquid spore suspension you 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.

Ethyl alcohol (70%) in spray bottle
Ethyl alcohol (70%) in 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.

Shroomok's Choice on Amazon:

Isopropyl Alcohol 70% USP Grade - One Quart Spray - $21.99

Isopropyl Alcohol 70% USP Grade - 4 X 1 Quart Spray Bottle - $45.00

6️⃣ Cotton pads — 2-3 pcs.

Shroomok's Choice on Amazon: Cotton Rounds Organic, 100 Count - $2.56

7️⃣ Grower's outfit: gloves, face mask, cap, medical gown (or PE raincoat).

Shroomok's Choice on Amazon: 10 Gloves and 1 Mask Bundle - $6.99

Grower's outfit
Grower's outfit: gloves, face mask, cap, medical gown (PE raincoat)

Preliminary preparation before making liquid spore syringe

1️⃣ Put on the necessary grower outfit: face mask, medical cap, gloves, medical gown (or PE raincoat).

2️⃣ Wipe the table with alcohol (by using alcohol in bottle or spray bottle and cotton pads).

3️⃣ Spray alcohol on your gloves. Wipe your hands well over the entire surface of the gloves.

4️⃣ Spray alcohol on the sleeves of the medical gown (or raincoat) up to the elbow.

5️⃣ 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.

6️⃣ 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.

Open the vial (ampoule) and fill in the syringe with sterile water for injections
Steps 1-2. Open the vial (ampoule) and fill in the syringe with sterile water for injections

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.

Liquid spore. No glovebox. Psilocybe cubensis growing
Steps 3-4. Don't open a zip bag! Squeeze all water from the syringe into the zip bag with spore print.

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.

Liquid spore syringe preparation in zip bag
Step 5. Rub spore print to separate spores and mix them 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.

Liquid spore preparation
Wrap liquid spore syringe in foil. You can see that water became black with mushroom spores

9️⃣ Sign the strain.

🔟 Leave prepared liquid spore syringe to rehydrate for 24 hours at room temperature (+20+25°С or 68-77°F).

Psilocybe mushrooms liquid spore suspension preparation for further inoculation substrate
Leave liquid spore for 24 hours for rehydration before substrate inoculation

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?

Ready-made liquid spore syringe could be stored up to 1 year and spores stay viable.

Liquid spore syringe should always be wrapped in foil (to avoid light) and stored in the refrigerator at +2°..+8°C or 35-46°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 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.

Make all manipulations for liquid spore syringe preparation in zip bag. Then simply add another 5-15 ml of water to fill in the whole volume of the syringe.

You can dilute the liquid spore till solution is slightly colored with spores.

While the spores are 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 ▶️

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