Description
Liquid Culture Expansion Kit Instructions
Please read these instructions carefully to avoid mistakes. Making a single mistake can ruin the whole kit. Please also watch our video.
About This Kit
We sell glass sterilised jars, but they are only available for pickup from our Marrickville store. If you don’t live nearby, this kit is the perfect fit for you.
This kit is designed for beginners with no prior experience in mycology or equipment. You only need a liquid culture syringe (either purchased from us or another supplier) and a clean and still workspace.
IMPORTANT:
This kit is designed to expand liquid culture by a factor of 10. Do you have one syringe from a supplier? Now with this, it’s 10 syringes in about 10 days. You cannot use it to expand spore syringes โ it is highly discouraged due to the risk of contamination.
Spore Syringe vs Liquid Culture
- A liquid culture (LC) syringe contains white, jellyfish-like mycelium suspended in nutrient liquid.
- A spore syringe contains spores, which appear as brown specks (visible or not). While expanding a spore syringe is technically possible, we strongly advise against it. Spores are not guaranteed to be sterile, so contamination is likely to occur.
Testing Your Culture
You should test your starting culture to ensure it’s clean.
Add one drop of your liquid culture to the MEA cup (included in the kit) and wait at least 5 days.
- If only white mycelium grows, your culture is clean.
- If any discolouration appears, itโs contaminated.
Whatโs Included in the Kit
| Item | 125 ml Jar | 250 ml Jar | 500 ml Jar | 1000 ml Jar |
|---|---|---|---|---|
| Sterile Nutrient Liquid Culture | 2x 50ml | 4x 50ml | 8x 50ml | 16x 50ml |
| Disposable Sterile Syringes | 11x 10ml | 21x 10ml | 21x 20ml | 41x 20ml |
| Disposable Sterile Needles (18G) | 25 | 45 | 45 | 85 |
| Syringe Caps | 10 | 20 | 20 | 40 |
| MEA Test Cup | 1 | 1 | 1 | 1 |
| Marble | 1 | 1 | 1 | 1 |
| Alcohol Spray Bottle | 1 | 1 | 1 | 1 |
1x Sterile jar with marbleโ ๏ธ Do not open this jar!, Do not open the jar as its pre sterilised.
MEA (Malt Extract Agar) test cup. We highly recommend to order 1 extra MEA cup, one to test the liquid culture from yiur supplier, and one to test the liquid culture once its expanded.
Before You Start
- Work in a clean environment
- Wash your hands
- Wear gloves and a mask (recommended)
- Use the alcohol spray to clean the injection port and any surfaces
Step-by-Step Instructions
1. Prepare the Sterile Jar
IMP-Do not open the jar
- Spray the black injection port on the jar with alcohol and wait 30 seconds.
- Inject all the nutrient liquid from the syringe we provided into the jar through the injection black port.
2. Inoculate the Jar
- Respray the injection port with alcohol. Wait 30 seconds.
- Inject the entire 10ml liquid culture syringe into the jar(That you got from us or another supplier). It may work with even 1ml, but the more starting culture, the better. 10ml is recommended as it provides a perfect expansion ratio. You can use more than 10ml, but avoid using less than 10ml.
- โ Do NOT mix two different syringes, even if they are the same strain.
- Label the jar with the strain and the date.
3. Incubate the Jar
- Leave the jar undisturbed.
- After 5โ10 days, gently shake the jar to distribute the mycelium. The marble helps break up clumps.
4. Test the Culture
- Spray the injection port with alcohol.
- Take a new sterile syringe and needle.
- Swirl the jar(don’t get the white filter wet), insert the needle through the black port, and draw 1 mL of culture.
- If you tilt the jar to draw the liquid, ensure the white filter does not touch the liquid; if it does, it’s not a problem, but try to avoid it as much as possible.
- Inoculate the MEA test cup with a few drops of the sample; do not add the entire 1 mL of liquid.
- Wait at least 5 days.
- If the plate stays white and clean, your expanded culture is safe. If the test did not yield clean results, there is a chance that no sterile technique was maintained or a contaminated starter culture was used. We highly recommend buying an extra Mea cup (not Meag) to test your starter liquid culture before using this kit.
- Dispose of the used syringe and needle safely โ never reuse them.
After Successful Expansion
- Pull liquid from the jar into new sterile syringes.
- Soak caps in isopropyl alcohol before placing them on filled syringes. Watch the video on how to do this.
- Label each syringe with the strain and date using masking tape (markers may rub off due to alcohol exposure).
- Store syringes in the fridge for 6โ12 months, depending on strain.
- Some cultures are cold-sensitive โ ask your supplier.
Storage and Reuse
- After using the jar, wash it thoroughly.
- Keep the injection port and air filter on the lid.
- Dry thoroughly and store in a zip-lock bag in a clean, dry place. Do not store in a ziplock bag if not 100% dry.
You may be able to make some more liquid culture in the microwave to reuse it. How to sterilise liquid culture in a microwave.
Our plastic jar, lid, injection port, air filter, and marbles are all microwave-safe (so basically everything).
Common Mistakes to Avoid
โ Reusing needles โ use a new sterile needle every time
โ Skipping the test for your starting liquid culture
โ Skipping the test of your expanded culture before use
โ Using contaminated liquid culture to inoculate grain bags
Timeline Summary
Option A โ Safer Route:
- Test your LC syringe on the MEA cup โ wait 5 days
- If clean, add to the jar and expand โ wait 10 days
- Test final jar culture โ wait 5 days
Total time: ~20 days
Option B โ Faster but Riskier Route:
- Skip testing, add the LC syringe directly to the jar, and wait 10 days
- Test final jar culture โ wait 5 days
Total time: ~15 days
This Kit Does NOT Include:
- Extra MEA cups – You may want to buy them to test the liquid culture you purchase.
- Gloves or masks
- Grain bag or all-in-one grow bag
If you’re an absolute beginner and feel unsure, we have a comprehensive instructional video available in the ‘Instructional Video’ tab. Watch us do it.
Stay clean, stay safe, and happy growing!
