How To Make Liquid Culture

Contents

How To Make Liquid Culture – Step By Step Guide

When growing mushrooms, a starter culture such as liquid culture is utilized as a starter culture. Either to inoculate agar plates or sterilized grain for the purpose of breeding or to extend and develop more liquid culture are the two main uses for it. It is common practice to make liquid culture in jars with modified lids from a sterilised broth with a specified sugar to water ratio in a modified lid. A mycelium culture is added to this soup to give it a mushroom flavor. The mycelium will absorb the sugar and continue to grow, resulting in virtually limitless quantities of mycelium being produced.

Sterilization of the needle point of a syringe can be accomplished using heat to avoid the spread of impurities.

How To Use Liquid Culture

When sterilised grain is inoculated into a liquid culture, it can be utilized to make grain spawn. The sterilised grains can be inoculated with liquid culture as low as 2 cc of well colonised liquid culture when the procedure is used aseptically. Mycelium will consume the grains as it colonizes them over the next 2 – 4 weeks, feeding on them.

Preparing Jars For Liquid Culture

Liquid culture requires fresh air exchange for healthy development. As mycelium grows it releases carbon dioxide. We must modify the lids in order to allow for more oxygen to be introduced. Each modified lid will include a filter for fresh air exchange as well as an injection port that will allow a syringe to be used to inject and extract material without contaminating the environment. If you prefer, you can purchase modified lids from this website. Learn more about making liquid culture jars in our step by step guide by following the link below.

It requires a sugar concentration of 4 percent .

We prefer light malt extract as powders are easy to measure.

Brewers yeast is an optional supplement for quicker growing mycelium.

Buy Modified Mason Jar Lids

The creation of mushroom grain spawn and liquid culture is made simple with the help of modified mason jar lids. With a filter patch for fresh air exchange and reusable injection apertures for simple inoculation with a liquid culture syringe, each lid is designed to be user-friendly.

Step 1. Measure Ingredients

Precision scales are used to quantify the light malt extract and the brewers yeast in the brew (optional). Each Mason jar should be half-filled with liquid culture broth. Add a piece of glass marble or an amagnetic stirrod to complete the look. This will aid in the breaking up of clumps of mycelium later on in the process. Place a customized lid on each mason jar and wrap it with a piece of aluminum foil to keep it from becoming dirty. Sterilize the liquid culture broth for 20 minutes at 15 PSI under pressure in a pressure canner.

The cooled sterile sugar broth can now be infected with a live mycelium culture, which will grow in the absence of oxygen.

Using a liquid culture syringe that has been flame sterilized and injected into the injection port is possible.

Every day, gently spin the liquid culture jar to provide enough oxygenation. It is important not to splash fluids near the lid, since this may allow undesired microorganisms to enter. The greatest results may be obtained by stirring the liquid culture using an amagnetic stirrer.

Magnetic StirrersBetter Liquid Culture

With the use of a magnetic stirrer, the liquid culture may be better and more thoroughly oxygenated, which will result in faster and more robust development over time. Using a magnetic stirrer, you may break up the mycelium into minute fragments, which will increase the number of inoculation points available for the best potential outcomes. During the first 14–21 days, a significant amount of mycelium will have developed in the liquid culture. Using this method, you may inoculate more liquid culture, agar plates, or even grain spawn into your system.

Liquid Culture Collection

Liquid culture is a very effective technique in the mushroom grower’s toolbox. One or two droplets of liquid are all that is required to initiate the metamorphosis from sterilised grains to strong mushroom grain spawns. The liquid culture can be used to inoculate agar plates, seed grains, or even to provide an unending supply of liquid culture, depending on the application.

How To Make Liquid Culture For Mushroom Growing

It is commonly referred to as “liquid culture” or “LC” since it is made up of mycelium floating in a nutritional broth. This product was created to make the process of inoculating substrates and grain spawn more convenient. The risk of contamination is reduced because the culture is transferred from a sterile environment (within the jar) to a sterile syringe and then injected into a new, sterile environment rather than being transferred with agar wedges, which increases the risk of contamination, particularly for less experienced growers.

  1. For this reason, working as cleanly as possible with your culture and LC is extremely important to avoid contamination.
  2. With semi-regular mixing and shaking, you should see that the mycelium has ‘clouded’ the culture and spread throughout it after 7 – 14 days of starting the culture.
  3. Incubate at the right temperature for your species; I’m using blue oysters, so I’ll incubate at roughly 24 degrees Celsius for the duration of the experiment.
  4. To avoid repeating the steps above, stir/swirl your culture and collect a sample by pulling it into an injection port of a heat-treated needle and depositing a single drop onto agar.
  5. You may continue to use your sample in the confidence that your LC is clean and ready to use if it is clear and colonizes normally without contamination.

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Mushroom Liquid Culture: The Easiest Guide to Using Liquid Culture[2022]

When growing mushrooms with liquid mushroom culture, you may find yourself feeling more like a scientist or mycologist than a mushroom enthusiast. Fortunately, you do not require a degree to effectively develop your own mushroom liquid culture, or to transform that culture into mycelium that will ultimately produce healthy mushrooms in your own garden or at your residence. If you think this technique sounds interesting, continue reading. By the time you’ve finished reading this tutorial, you’ll be ready to begin.

What is Mushroom Liquid Culture?

Liquid Culture is a sterile combination of water and one of a few particular sugars that is used to cultivate bacteria. (We will explain how to produce liquid culture in the following section.) Once the mushroom spores have been injected into the soil, the goal of this nutritional combination is to promote the growth of mycelium. Imagine mycelium floating in a nutritional soup, and that’s what liquid culture looks like. Liquid culture simplifies the process of inoculating substrates. Having established itself in the nutrient-rich sugary broth, the mycelium-rich combination may be inoculated onto a substrate of your choosing or preserved as a live mushroom culture.

What are the advantages of using Mushroom Liquid Culture?

  • The possibility of contamination is reduced
  • In the opinion of Liquid Culture Shroomery, the use of liquid culture shortens the incubation period. Mycelium development is virtually limitless once it begins
  • Once mycelium begins to develop, it continues indefinitely.

What are the disadvantages of using Mushroom Liquid Culture?

  • It is critical to maintain a sanitary working environment. Improved suitability for advanced mushroom cultivation
  • In liquid culture, there is no evidence of contamination. When you utilize your concoction, you will only be aware that it has been tainted.

A suspension of mycelium in a liquid culture of mushrooms

How to Make Mushroom Liquid Culture

This recipe may be used to learn how to manufacture Oyster mushroom liquid culture if you are interested in learning how to do so. In fact, this recipe may be used with whatever type of mushroom you want to use.

What you will need:

  • Jar used to store grain, often known as an airport jar
  • A pressure cooker
  • Syringes (10mL each)
  • Needles (Note: the larger the number, the smaller the needle). a pressure cooker We recommend that you use 18 gauge). To combine with water, one of the following sugars should be used: Organic honey, corn syrup, corn sugar, light malt extract, and dextrose (glucose) are among the ingredients.

It is not recommended to use sucrose (household sugar). Honey was our selection for this guide. What is the optimal sugar-to-water proportion? A 4 percent ratio is considered optimal. This amounts to 4cc, 4ml, or 1 teaspoon of honey per 100ml of water, depending on the measurement. However, excessive sugar (more than 10 percent) is hazardous to mycelium, so keep this ratio between 3 and 5 percent in mind.

A step-by-step guide to making your own mushroom liquid culture:

  1. Use eco-friendly measuring cups to carefully measure and combine the sugar and water in a jar before freezing. Warm water will aid in the rapid dissolution of the sugar, although it is not required. Close the jar with its lid
  2. Aluminum foil should be placed over the top of the jar before placing it in a pressure cooker. Cook at 15 psi for no more than 15-20 minutes at maximum pressure. Excessive heating of the sugar will result in caramelization and, as a result, poor to non-existent mycelium development. Allowing the pressure cooker to cool naturally before removing the container is highly recommended. Take precautions. The jar will be quite hot when it is opened. Allow for the jar and its contents to come to room temperature before using. Incorporate a mother culture into your liquid culture (see below for information on where to get mother culture)

Important Note: If you are not using an airport jar, wait until you are ready to inoculate your substrate before removing the lid and replacing it as soon as you are finished. This will prevent airborne pollutants from entering the jar and destroying your mycelium.

How to add mother culture to your liquid culture

Following their time in the pressure cooker, you may add a few ccs of mother culture to each of the jars once they have completely cooled. Shake the mother culture vigorously to dislodge any mycelium that has formed. Hands should be properly washed with an environmentally friendly soap and dried with a clean paper towel. If you are using a syringe needle for the first time, be sure it is sterile before you begin. If not, sterilize the needle by holding it in the flame of a lighter or a candle until it is red hot, then discard the needle.

To cool the needle down, clean it with an alcohol swab.

If you have more than one jar, repeat the process. Cleaning the outside of the jars with biodegradable wet wipes is a good idea before using them.

What to do with a jar of mushroom liquid culture

Simply place it on a shelf and wait for it to sprout leaves and grow. The ideal temperature for most mushroom cultures is approximately 77°F (25°C). Any difference in this temperature will cause them to grow at a slower rate than normal. For 4 to 5 days, do not open or close the jars. You will observe the mycelium expand as it takes the sugar during this period because it is in its development stage at this point. The culture should be swirled every couple of days after it has been resting for a few days to break up the mycelium and provide oxygen into the culture.

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When you are ready to inoculate your substrate, transfer the culture to agar, or share some of the culture with your friends, tilt the jar so that the liquid reaches the injection port and, using a sterile syringe and needle, suck the culture into the syringe until it is completely filled with the culture.

How to Use Liquid Culture to Grow the Perfect Mushrooms

The process of going from having liquid mushroom culture to harvesting a crop of edible mushrooms is a little more involved. The first stage is to increase the size of the mycelium. This may be accomplished by employing a variety of grains, including rye, wheat, oat, birdseed, and corn seeds, among others. You might also use cardboard or wood chips as an alternative (for this guide, we chose grain).

Prepare the grain to receive the liquid culture

  1. Soak the grain for 12 to 24 hours before boiling it to lessen the amount of time it takes to cook. Cook for 10 minutes if the beans have been soaked, or 20 minutes if they have not. Ideally, the grain should be boiled for a long enough period of time to become hydrated and soft, but not mushy. Grain that has been overcooked is more likely to deteriorate. Remember that your grain will be cooked in a pressure cooker once more later on, therefore it is preferable to have it slightly undercooked at this point. Practical Tip: Some individuals skip the boiling stage and proceed directly from soaking to bagging and then placing it in the pressure cooker without completing the process. Colander the grain, then tip it onto a table covered with kitchen towel to drain. Allow it to dry completely before using. It is possible to speed up the process by using a bamboo charcoal dehumidifier and a fan. If the grain is too warm or wet when it is placed in the bag, it will get contaminated
  2. Otherwise, it will not work.

Put the grain in a bag and place it in a pressure cooker

  • Bags should be covered with aluminum foil to prevent them from melting if they come into touch with the pot’s edge. A melted bag, as well as the grain contained within it, must be discarded. Cook the bag of grain for 90 minutes at 15 PSI in order to disinfect the grains.

Add the liquid culture

  1. Allow the bag of grain to cool before taping it shut using masking tape. Take care not to obstruct the filter patch, since this is necessary for air exchange, which is necessary for the growth of the mycelium. Your bags are now ready to be infected with your liquid mushroom culture, which you may do at this point. In other circumstances, already infected grain or agar may be used instead. Simply inject your liquid culture directly into the bag using the injection port, and the bag will be filled. Bags without an injection port should be used if possible
  2. If this is not possible, inject through the tape and cover the needle-prick hole with another piece of tape.

Wait for your mycelium to grow

  1. Place your bag in an upright posture in a quiet area where it will not be disturbed and where the temperature is between 75 and 81 degrees Fahrenheit (24 and 27 degrees Celsius). Make certain that there is sufficient space between the filter, the plastic, and the grain to enable for air exchange to take place between the components. It makes no difference whether it is bright or dark. You should begin to observe evidence of development 3 to 10 days following the inoculation procedure. By the end of 3 or 4 weeks, your mycelium should have colonized 70 to 75 percent of the bag’s surface area. If you don’t do it at this time (or sooner, you risk postponing the development), you’ll have to use your hands to break up the grain as it passes through the bag in order to speed up the last step of colonization. Three to seven days after the mycelium has been split up and spread throughout the bag, it will have colonized the whole bag.

Spawn the colonized grain to a bulk substrate

  • In order to choose the bulk substrate to employ, you must first determine the sort of mushrooms that will be grown in your liquid culture. Among these are theAgaricus bisporusfamily (White Buttons, Cremini, and Portobello), oyster mushrooms (which prefer to grow on anything made of wood), and straw mushrooms (which grow on straw, as their name indicates
  • Learn more about cultivating straw mushrooms here).

Tend to your mushrooms

  1. According on the sort of mushroom you select to cultivate, you will need to feed and care for it until it is ready for harvest. The growth of your own mushrooms is covered in a number of articles on our website, so whether you’re looking for information on which mushrooms are the simplest to grow, which ones are the most nutritious, or even where to get mushroom kits, you’ll find it all here

8 Best Places to Buy Liquid Mushroom Culture

There are a variety of websites where you may get liquid culture:

1.True Leaf Market

True Leaf Market has a wide selection of high-quality, organic seeds, mushroom cultures, and growing kits in addition to other gardening supplies. Their company, which was founded in 1974, has been servicing home and commercial growers with amazing success. There is a large selection of mushroom types available on their website, as well as thorough manuals and kits. True Leaf Market goods are organically certified and free of genetically modified organisms. Each mushroom growing kit includes comprehensive instructions that make it simple to produce flush after flush of high-quality mushrooms in the comfort of your own home.

The following items are included in the kit:

  • A sawdust block that has been completely sterilized and colonized with a robust commercial mushroom strain
  • A small mister, a humidity tent, and detailed growing instructions are included.

Price: Starting at $27.99Website:

2.Mushrooms.com

Mushrooms.com has a large variety of gourmet, edible mushroom liquid cultures and spore prints in a variety of flavors. They also carry a variety of therapeutic mushroom species. Their concentration is on cultivating species that are difficult to locate for sale and that are useful to have on hand for your own personal use.

Their kits contain spore culture syringes, which are easy to use and are great for novices, as well as spore prints, which are more advanced and are suitable for more experienced growers as well. The following items are included in the kit:

Price ranges from $17.95 to $19.50Website address:

3.New Generation Mushroom Supplies

Based in Melbourne, Australia, New Generation Mushroom Supplies is an online mushroom growing kit and supply company that specializes in mushroom growing kits and supplies. Their product line includes mushroom spores, liquid culture syringes, equipment, and other related items. You may save time and money by shopping at this location because they offer everything you need, including the liquid mushroom culture, in one convenient location. The following items are included in the kit: Price ranges from $38.50 to $40.70.

tax and shipping) Website:

4.Fungi Ally

Fungi Ally distributes pure, certified organic mushroom goods such as spawn and grow kits, as well as mushroom growing supplies. Their spawn is available in three different varieties: sawdust spawn, plug spawn, and grain spawn. Their kits are a ten-pound block that has been fully colonized and is ready to be planted. The Fungi Ally website also offers a wide range of tools and equipment, in addition to mushroom extracts and other mushroom-related items. They also have cultivation manuals and workshops available, so have a look at their website for some pretty useful materials and information!

  • Grow Kits: 1 x colonized 10 pound block (which produces 3-5 pounds of mushrooms)
  • Grain Spawn: 1 x 6 pound bag of grain spawn
  • Grain Spawn: 1 x 6 pound bag of grain spawn Sprouted Sawdust Spawn: one (1) bag of sawdust spawn
  • Sprouted Plug Spawn: one (1) bag of spawn plugs

Price:Varies depending on the product – please see their website for additional information. To obtain a 15% discount, visit the website and use the promo code mindseteco.

5.Root Mushroom Farm

Root mushroom farm cultivates and delivers liquid cultures that are simple to produce and yield results in a short amount of time. Their goods are completely natural and environmentally friendly. In their opinion, raising mushrooms at home with the help of their liquid cultures should be a lot of fun for everyone involved. They guarantee complete customer happiness, or they will refund your money. Liquid culture from a mushroom farm’s roots. The following items are included in the kit: Price:$13,99Website:

6.Mushroom Hobby

Mushroom Hobby is passionate about sharing their mushroom-knowledge with mushroom enthusiasts from all over the world, and they welcome visitors from all over the world. On their website, they also provide blogs and recipes that they have created. You may learn everything you need to know about grain spawn, mycelium, agar dishes, and more. Make sure to check out their website for delectable dishes that use the mushrooms you’ve just harvested! The following items are included in the kit:

  • 10-12 CC of liquid culture
  • 1 new needle
  • 1 alcohol wipe
  • 1 syringe loaded with liquid culture (10-12 CC)
  • A one-page instruction manual on how to use the syringe

Price:$15,00 Website:

7.Maya Mushrooms

Maya Mushrooms cultivates edible, specialized, and medicinal mushrooms, as well as grow-at-home kits and growing materials for customers worldwide. They believe on offering high-quality, locally grown, nutrient-dense vegetables at a fair price to their customers and community. They aspire to be carbon neutral while also working in symbiotic partnership with nature by growing their mushrooms from up-cycled waste product and using it as fertilizer. Maya Mushrooms are a liquid culture that grows mushrooms.

  • (1) 10cc syringe of liquid culture
  • (1) 18 gauge needle that has been sterilized
  • (1) Alcohol wipe

Prices start at £10 and go up from there.

8.Liquid Fungi

Liquid Fungi’s cultures are obtained in a laboratory from the best samples available anywhere in the globe. The company tests and guarantees that each culture is 100 percent contaminant-free, or they will replace it. For optimal yield and minimal incubation time, each syringe includes a proven, high-output, isolated sub-strain. It’s called Liquid Fungi, and it’s a liquid mushroom culture. The following items are included in the kit: Price:$10,49Website:

Conclusion and Our Favorites

Because of the large number of cultures available from Liquid Fungi, they are our preferred source by far. The company’s website does not specify if their kits contain a needle and an alcohol wipe, which is disappointing. They do, however, give thorough environmental information for each variety, allowing you to know ahead of time what growth conditions you should be looking for before placing your purchase. To get started, you may get mushroom liquid culture from any reliable provider, and your mushroom-growing adventures will be off to a fantastic start.

Frequently Asked Questions

No, all mushroom liquid culture recipes are identical in their preparation. The type of mushrooms you harvest will be determined on the mother culture you utilize.

What sugar can I use in my mushroom liquid culture recipe?

Organic honey, corn syrup, corn sugar, light malt extract, and dextrose are all acceptable substitutes (glucose). Do not use ordinary table sugar (Sucrose).

What is the disadvantage of liquid culture over agar?

When opposed to a plate of agar, where contamination may be seen immediately, contamination in liquid culture is difficult to detect. When you utilize your concoction, you will only be aware that it has been tainted.

References

Photographs courtesy of Getty Images /ossyugioh The majority of LC recipes are designed to have a sugar level of no more than 4 percent by weight of the finished product. More than this is harmful to mycelium’s growth. Other elements, such as protein, fats, carbohydrates, minerals, vitamins, and other nutrients, can be included in modest amounts. Adding a little amount of an infusion or a few particles of the final substrate during the spawn run can help to accelerate myceliation. Keep in mind that some components might induce cloudiness, which can mask the presence of contamination.

When developing your cultures, be sure to switch up the recipe to keep things exciting for your cultures.

  1. The Getty Images collection contains a variety of images that are available for licensing. /ossyugioh In most LC recipes, the sugar level is 4 percent or less by weight, according to the manufacturer’s specifications. It is poisonous to mycelium if you consume more than this. Other substances, such as protein, fats, carbohydrates, minerals, vitamins, and other nutrients, can be added in modest amounts to enhance the nutritional value. Myceliation may be accelerated during spawn run by adding a little amount of infusion or a few particles of the ultimate substrate. Keep in mind that some components might induce cloudiness, which can make contamination difficult to detect and distinguish. Mason jars with a regular opening (500 mL) are my go-to container for most things. When growing your civilizations, switch up the recipe a little to keep things exciting for your people. For this recipe, you will need the following items: a digital scale or measuring spoons
  2. A measuring cup
  3. Jars
  4. Airport lids and rings
  5. Agitators (marbles
  6. Pieces of broken glass
  7. Crystals
  8. Or magnetic stir bars)
  9. An electric pressure canner (PC) or an alternative
  10. Aluminum foil
  11. And a pressure canner (PC).

Tissue Culturing to Liquid Culture

Image courtesy of Isabelle Kirouac By stabbing a mushroom, you can take a sample of tissue. Generally speaking, this biopsy approach works well with most fleshy mushrooms, while tough or small mushrooms might be difficult to collect with this method. Mushrooms that have been waterlogged should be allowed to dry out in the refrigerator first. Dry mushrooms can be rehydrated by soaking them in a solution of 3 percent hydrogen peroxide. Repeat the technique with additional jars of media to increase your chances of success.

  1. Isabelle Kirouac contributed to this photograph. Mushroom stabbing is used to take a sample of tissue. However, thick or microscopic mushrooms might be practically hard to collect with this approach. This biopsy technique works well with most meaty mushrooms. Mushrooms that have been waterlogged should be allowed to dry out in the refrigerator before cooking. In order to rehydrate dried mushrooms, they should be immersed in 3 percent hydrogen peroxide for 30 minutes. Using many jars of media will increase your chances of success. You’ll need the following supplies: a mushroom, a sterile syringe fitted with a 16-gauge needle, a sterilized jar of water with an airport lid, a sterilized jar of LC medium with an airport lid, alcohol in a spray bottle, alcohol prep pads, cotton balls, tweezers or a knife (optional), and a permanent marker for marking your results.
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Making a Spore Syringe

When a spore syringe is transferred to LC, grain, or substrate, the result will be multiple spore germination, which will result in several genets growing together in the same space. Selections of fruits from this lineage can be tissue grown in order to identify a suitable strain for further study. Image courtesy of Isabelle Kirouac Preparing to move liquid culture from one location to another. Supplies required include a spore print bag (see “Spore Prints” above), a sterilized water jar with an airport cover, cotton balls, a sterile syringe, alcohol spray, and an alcohol flame (or a replacement for an alcohol flame).

  1. Assemble items on a clean work surface and wash your hands well
  2. Unwrap the syringe, but do not remove the cap just yet. Remove the foil from the jar, spray the SHIP with alcohol, and wipe the jar clean with a cotton ball
  3. Apply one more spray of alcohol to the SHIP of the water jar to finish it off. Remove the top from the syringe and swiftly enter the injection port
  4. Fill the jar with water until the needle tip is submerged, being careful not to wet the filter. Draw up a syringeful of water in your hand. For the time being, leave the needle in the port. Remove the needle from the jar and sanitize the exterior of the spore printing bag. With the bag on the work surface, pierce the bag (being cautious not to go through both layers) and inject water into the bag. To combine spores and water, use your hand to stir the mixture, then suck the water back into the syringe. Alternatively, if you want to produce many syringes, spritz the water jar’s SHIP with alcohol and inject spore solution into it
  5. Alternatively, if you want to make several syringes, spritz the water jar’s SHIP with alcohol and inject spore solution into it. Before utilizing the spores, label them and allow them to hydrate for 24 hours. Spore syringes can remain viable for several months to even years. Store in a cool, dry location away from direct sunlight.

LC or Spore Syringe to LC Transfer

It is necessary to have the following items: spore/LC syringe and alcohol flame (or an additional sterile needle), OR sterile syringe and an LC media jar, as well as alcohol spray and cotton balls. If you are using a pre-made spore or an LC syringe, you may skip steps 5 and 6 and go straight to step 7. Assemble all of your equipment and supplies on a clean work table.

  1. Unwrap the syringe, but do not remove the cap just yet. Stir the contents of both jars vigorously for approximately 30 seconds, breaking up the mycelium in the LC. Remove the foil from the LC media jar, spray the injection ports of both lids with rubbing alcohol, and wipe them clean with a cotton ball. Apply one more spritz of alcohol to the SHIP of the LC jar to finish it off. Remove the top from the syringe and swiftly enter the injection port
  2. Tip the jar until the needle tip is in the LC, taking care not to wet the filter in the process. This may be done either in midair or on a work surface if necessary. Preparing 1-10 mL of LC is important
  3. Make sure you get some mycelium in there, not just broth. If the needle becomes clogged, push a small amount of needle out to clean it and continue. Place the jar on its side
  4. If you’re using a pre-made LC or spore syringe, either replace out the needle for a sterile one or heat the needle until it’s red hot to sterilize it. Allow 3-5 seconds for the needle to cool before continuing. Spritz the SHIP of the media jar with alcohol and enter the needle into the media jar as rapidly as possible. Fill the jar halfway with the contents of the syringe. Label the jars with the species, strain, date, medium, source jar, and generation, and place them in the incubator to start growing

Agar Plate or Slant to LC Transfer

Scratching mycelium from agar into water.You’ll need the following supplies: an aseptic transfer space (also known as a still air box), a sterile needle and syringe, a sterilized jar of water fitted with an airport lid, a mycelial culture on agar (in either a Petri dish or a culture tube), an alcohol spray, cotton balls, plate wrapping material, and a permanent marker.

  1. Clean SA B or another aseptic transfer room should be used to store all supplies. Mist the interior of the box with water and let the air to settle
  2. Unwrap the syringe, but do not remove the cap just yet.

More fromDIY Mushroom Cultivation:

  • How to Make a Home Fruiting Chamber
  • What is a Mushroom
  • How to Grow a Home Fruiting Chamber

This is an excerpt from Willoughby Arvalo’s book, DIY Mushroom Cultivation. Willoughby Arvalo’s book, published by New Society Publishers in 2019, is a must-read. All intellectual property rights are retained.

Select Liquid Media Recipes (Per 1000 ML Water)

Consider these recipes as jumping-off points, and then let your imagination go wild. Keep sucrose to a minimum (cane or beet sugar). Complex carbohydrates have the potential to promote more rapid development than simple sugars alone. A pinch of gypsum and/or nutritional yeast can be added to any of these dishes to boost the amount of minerals and vitamins they contain, but this will lower the clarity of the dish. It is more accurate to measure by weight than than by volume. Fastfred’s Media Cookbook has further information.

Malt Extract LC (MELC)

  • 4 tbsp (40 grams) light malt extract
  • Malt Extract Dextrose LC (MDLC)
  • 2 tbsp (20 grams) light malt extract
  • 2 tbsp (20 grams) dextrose
  • Honey LC (HLC)
  • 2 tbsp (40 grams) honey
  • 2 tbsp (40 grams) dextrose
  • 2 tbs

Sabouraud’s Dextrose Broth (SabDex)

  • 4-tablespoon dextrose (40 grams)
  • 1-tablespoon (10 grams) polypeptone (neopeptone)
  • 1-liter pure water Complete LC (CLC) is an abbreviation for Complete LC. light malt extract (about 20 grams) 2 tablespoons 2 grams peptone
  • 0.6 grams yeast
  • 10 drops vegetable oil
  • 2 grams ground grain
  • 2 grams ground wheat

Grain Cooking Water LC (GCWLC)

Whole grain water, either full strength or diluted, that has been kept from the boiling process (i.e., for grain spawn). Use immediately or freeze immediately for later use. Pour through a coffee filter to remove any impurities (but some batches will remain cloudy). Because grains contain numerous robust bacterial endospores and because there are few simple sugars to caramelize, they should be sterilized at the same time as the grain, or for 1-2 hours. The kind of progress I achieve in this medium appeals to me.

Published on Aug 14, 2019

Learn about the strange and wonderful world of Mushrooms, including information on how they’re defined, what they’re made of, and how they’re classified.Copyright 2022, All Rights Reserved | Ogden Publications, Inc. Select and build the right fruiting chamber to start your own home mushroom cultivation project using one of these chamber methods.

How to Make a Liquid Culture for Mushroom Cultivation

Liquid Culture is a technique for increasing the rate of colonization of your grain spawn. A nutrition supply might awaken latent contaminants/endospores in grains that are poised to take advantage of the opportunity. Don’t let them get away with it! Use liquid culture to defeat them before they get a chance to get to the food. * In this article, the term “liquid culture” refers to a sugar-based solution rather than spores in a liquid syringe, which will be defined as follows: The cleanliness of your culture and grain spawn will have a direct impact on the success of your LC.

2) Perform 2-3 agar transfers to obtain a monoculture before transferring it to your LC.

Agar Jar is available for purchase right now. Getting Things Started Sugar is a favorite of mycelium, thus sugar will be the primary element in your liquid culture. There are a variety of approaches that may be used to do this, including Karo, Light Malt Extract, Honey, and others.

  1. This will be accomplished through the use of Light Malt Extract powder. This may be purchased at a brewery store or via an internet retailer such as Amazon.com. Fill your 8oz jar halfway with about 1/8th of a teaspoon. * Please keep in mind that a filter patch or syringe filter must be placed on the top of the jar in order to allow for some air exchange. Fill the jar halfway with water. Fill it to the point where there is approximately a half inch of gap between the lid and the liquid
  2. Close the jar’s lid. Closed liquid culture jars should be sterilized for about 15 minutes at 15 psi. Allow time for cooling.

Phase of Inoculation Once the liquid culture has cooled, you’ll be able to inject some culture into agar wedges or place them in an agar plate. It will take around one week until your culture is ready to be used with your grains. * You may get all of these components in my store, if you don’t want to construct your own jars or liquid culture from scratch.

How to make liquid culture? – Instruction Guide

Advanced mushroom cultivation may be a thrilling and rewarding experience. It is likely that you enjoy learning about approaches that can assist you in customizing your growing procedure and optimizing the output of your magic mushroom harvest. One of these approaches is learning how to utilize and manufacture liquid culture, which is one of the techniques. Have you previously benefited from the countless advantages of this yellowish-brownish liquid? It has the potential to be more cost-effective and to provide outcomes more quickly than in the past.

Even better, after reading this essay, you will be able to build your own liquid culture!

Let’s have a look and see!

  • What is Liquid Culture
  • When should Liquid Culture be used
  • How should Liquid Culture be made

What is Liquid Culture?

Liquid Culture is made out of a nourishing solution that has been sterilized. This is often a blend of water and a variety of sugars of various sorts. When spores are introduced into the mixture, it has the capacity to produce mycelium, which may be used to create mushrooms. The advantages are as follows:

  • Faster results: Because the cultivator no longer has to wait for spores to germinate, incubation durations are reduced by at least a week
  • Faster results Mycelium growth is virtually limitless: a single cc of spore solution may be used to produce an infinite volume of liquid culture. Because once mycelium begins to grow, it continues to grow indefinitely

Make careful to follow these step-by-step guidelines to ensure a smooth transition through the procedure. This post will concentrate on the process of creating Liquid Culture. Get up to speed with this instructive essay about what Liquid Culture is all about.

When to use Liquid Culture?

Liquid culture is ideal for advanced mushroom production since it is cost effective. Starting from the beginning with PF Tek? Make certain that the mushroom spores are injected into the liquid culture before preparing the substrate! What if you want to take it easy with theGrow Kits sans Mycelium? This is an excellent pick! Because these Grow Kits sans Mycelium include a substrate that is ready to use as well as a liquid culture vial, they are a great value. Is there an alternative to Mycelium Growkits?

How to make liquid culture? – Honey/Water Instructions

A 4 percent dilute solution of different sugars and other nutrition sources in water is typically used in liquid cultures to maintain their viability. For example, 96 mL/cc water has 4 grams of sugars per 96 mL/cc water. (Water weighs one gram per milliliter/cubic centimeter.) The most often utilized nutrition sources are as follows:

  • Honey (while non-organic honey has been reported to be effective, organic honey is strongly recommended.)
  • Karo, corn sugar, light malt extract, and Dextrose (glucose) are all ingredients in this recipe.

It is not recommended to use regular table sugar (sucrose).

In order to make this instruction guide as simple and accessible as possible, we would want to concentrate on the nutritional source that is most widely available: honey!

How much honey?

It is necessary to have a 4 cc/ml concentration to achieve the desired 4 percent ratio. Alternatively, 1 teaspoon can be used to measure 4cc/ml of honey. One teaspoon of organic yellow honey to one hundred milliliters of water is a close approximation to the correct ratio! Don’t be concerned if your solution is a bit off (3 percent to 5 percent). It’s important to remember that it’s preferable to be too weak rather than too strong, because too much sugar (about 10%) is poisonous to the mycelium and will prevent anything from growing there.

Mixingcooking

Once you’ve decided on a method, it’s time to get to work mixing things. It is possible to heat or reheat the water before adding the sugars in order to speed up the dissolution process. The top of the jar should be wrapped with aluminum foil before it is placed in the pressure cooker. Slowly increase the pressure to 15 psi over the course of 15-20 minutes. That is no longer the case! When working with honey, it’s crucial to remember that if you pressure cook for an excessive amount of time, your solution may become yellow.

Allow the pressure cooker to cool completely before removing it from the heat.

How to make a vacu-tainer?

  • Make a tiny hole in the lid/screw top of the jar that is large enough to accommodate a syringe needle. It’s recommended to use half pints for this. Using a glob of silicone sealant (preferably transparent), seal the edges on both sides. Wiggle it around until it is a centimeter thick around the hole on both sides.
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To begin, drill a tiny hole in the lid/screw top of the jar that is large enough to fit a syringe needle. This is best done with half-pints of beer. Seal it on both sides with a glob of silicone sealant (preferably transparent). Swirl it around to ensure that it is a centimeter thick around the hole on either side; and

Liquid Culture storage

Liquid Cultures can be kept in the refrigerator for up to 6-8 months (or longer). Some people choose to add a small amount of H2O2 (about 1-3cc) at this time since the mycelium is capable of handling it, and this can help avoid contamination. How is liquid culture created? With the help of the Liquid Culture User Guide, you may begin colonizing right immediately!

Measurement notes

  • One milliliter of water weighs one gram, while one tablespoon of dextrose weighs around ten grams. (1 tablespoon light malt extract weighs approximately 10 grams
  • Weights may vary somewhat). (10 mL honey weighs 14 grams
  • 1 tsp (5 mL) honey weighs 7 grams
  • 1 tbsp (15 mL) honey weighs 21 grams
  • 10 mL honey weighs 14 grams

Ready to use Liquid Culture vial

Are you looking for something that is user-friendly, sanitary, and ready to use? We recommend that you purchase them from our Magic Mushroom Shop. Liquid cultures are prepared under sterile conditions and packaged in convenient containers for transportation. Everything is ready to go! Make use of your own vial to experiment with maximizing mycelium growth!

Sources

Maintaining your own mushroom culture collection is both a time-saving and a gratifying experience for mushroom enthusiasts. It takes some practice to get the hang of it, but there are techniques that don’t require a lot of expensive equipment to get the job done. The pressure cooker will be the most important piece of equipment you’ll need. For the most part, individuals utilize one of two types of media to store their cultures: agar or liquid cultures. Liquid cultures have a negative reputation in the mushroom world, mostly because it is more difficult to detect contamination in liquid cultures.

However, by following a few easy procedures, you may ensure that your liquid cultures remain free of contaminants even when you are not operating in a sterile environment.

Working with Liquid Cultures

What you’ll need is the following:

  • The following items are required: a pressure cooker capable of producing 15PSI, jars with an airport top, a syringe and needle (ideally 16+ gauge), alcohol wipes or alcohol spray, a lighter, gloves

The quickest and most straightforward method of creating your own liquid cultures is to begin with a clean culture from a reliable provider and then extend it into multiple liquid cultures. The procedures to be followed are as follows:

  • Fill your airport jars with a liquid culture formula, which is commonly composed of tap water plus some sugar or nutrients
  • And To sterilise the liquid culture in the airport jars, pressure cook them for 10 minutes. A tiny amount of master culture should be added to each batch of sterilised liquid culture. Continue to wait till the culture has grown out before using it to inoculate grain spawn or your fruiting substrate.
Adding liquid culture to airport jars

This stage is extremely straightforward; the only element that requires a little trial and error is determining which formula would work best for the particular mushroom that you are attempting to grow. A simple recipe for oyster mushrooms and shiitake mushrooms is tap water mixed with maple syrup. Simply combine 1 teaspoon pure maple syrup with 1 cup boiling water, whisk well, and pour into jars with the lids put on top (see recipe below). I try to fill my liquid culture jars as close to the top as possible, allowing around 5cm (2 inches) of space at the top for air.

Another suggestion is to include something in your jars that will help to break up the mycelium.

Adding this will assist you in breaking up huge clumps of mycelium in order to help your culture develop more quickly and prevent it from clogging the needle when you suck it into a syringe while you are growing it.

This prevents the filter from being overly moist, and it also ensures that the jars are protected from contamination until you are ready to remove the alfoil from the filter.

Pressure cook the liquid culture

Add the jars to your pressure cooker and bring them up to 15 PSI for around 15-20 minutes on high for about 20 minutes. Follow the manufacturer’s directions for using your pressure cooker and always remember to fill it with water to the level of the base plate. Allow the culture to cool to room temperature, which will most likely take many hours; I normally leave it to cool overnight to ensure that it is completely cold.

Add some mother culture to the liquid culture

After you’ve allowed your jars to cool, prepare your master culture, alcohol wipes, and lighter by getting them all ready. Shake your master culture hard to dislodge as much mycelium as possible from the culture. If this is the first time you’ve used the syringe needle, you won’t need to use a lighter because the needle will have been sterilized before use. Removing the alfoil from the top of your first jar will allow you to insert a syringe into the self-healing injection port and pump a tiny volume of liquid culture (1-2 mLs) into the jar.

Liquid culture has been pressure cooked, aluminium foil on top to keep sterile.

Regardless of whether you’re using alfoil, it’s a good idea to wash the injection port with an alcohol swab before inserting a needle into it. Alternatively, you may use your alcohol spray (70 percent alcohol/methylated spirits, 30 percent water) to saturate the surface. Using a previously used needle for your syringe will need you to disinfect the needle before using it again. All that is required is that you connect the needle to the syringe and hold it over a flame, moving it back and forth, until the needle becomes red hot.

After that, you may pump a tiny quantity of culture out of the needle to cool it down, or you can clean the needle with an alcohol wipe to chill it down even more.

Airport lids ready for innoculation with liquid culture

The majority of mushrooms grow best at temperatures about 25 degrees Celsius; if the temperature is higher or lower than this, the mushrooms will develop more slowly. In order to allow the mycelium to get into development mode, I normally put my jars on a shelf indoors for the first 4-5 days without disturbing them. As the mycelium consumes the sugar, you should notice a modest but steady increase in its size. After the resting time, spin the culture every few days to ensure that oxygen is introduced into the liquid and that the mycelium is broken up.

Then tilt the jar so that the liquid culture comes up against the injection port and, using a clean sterile syringe and needle, suck as much liquid culture into the syringe as possible.

Working with agar cultures

What you’ll need is the following:

  • A pressure cooker capable of producing 15 PSI of pressure
  • Agar agar
  • Some nutrients for the agar petri dishes or other small containers made of glass or plastic
  • A scalpel or a dental pick

Agar is a jellylike material generated from seaweed; the Malay term for jelly is agar-agar, which is why you’ll hear it referred to as agar or agar-agar. Agar is also referred to as agar-agar in some circles. Either powdered agar by itself or with additional nutrients and maybe antibiotics are available for purchase. You can get agar at your local Chinese grocery for the least price if you’re just starting out, or you can purchase a little packet for a few dollars on eBay if you’re a more experienced cook.

In order to produce your own, you may just purchase malt extract separately and combine it with agar.

Drugs such as antibiotics are beneficial because they help to reduce the amount of bacterial contaminations that occur.

It is possible to use a tiny amount of hydrogen peroxide (purchased from your local chemist) to reduce the number of bacterial troubles you encounter if you do not want to use antibiotics.

Containers

As long as the container is sealable and either sterile or able to endure the high temperatures of a pressure cooker, you may use agar in any application you can think of. Petri dishes are the most frequent type of vessel, but if you want to keep things simple, you may repurpose tiny plastic takeout containers (made of polypropylene) or small glass jars instead. If you want to keep mushrooms for a longer period of time, you can make slants in test tubes.

Mixing and Sterilising

For the agar combination, you’ll need to combine some boiling water with agar powder before pouring it into your container. The consistency of the gel will be determined by the ratio of agar to water you use. Using 25g of agar per 500ml of water (about 5 percent), Paul Stamets suggests, although you may get away with using less. Agar is typically prepared in a single pot and then transferred to petri dishes after being pressure cooked. To use jars, you can mix the agar, then pour it into the jars before placing the jars on top of the pressure cooker to cook.

Combine the agar and boiling water in a mixing bowl, then pour it into the beer bottle using a funnel.

If you’re pouring into petri dishes, a polypropylene jug is the most convenient vessel from which to pour.

Using a metal scalpel to do culture transfers is not recommended, but wrapping it in aluminum foil and placing it in the pressure cooker with the container will ensure that it is thoroughly sterilized.

Pouring

Following pressure cooking, you must pour the agar into the jug while it is still hot, otherwise the agar will solidify in the jug and become unusable. You will, however, be more likely to burn yourself if you attempt to pour it into petri dishes while the water is still hot. You will also notice that the condensation on the petri dishes will be far more. It is customary for me to wait an hour after turning off the heat in my pressure cooker before opening it up and putting the agar into plates.

If you’re just starting out, this means utilizing a still air box or a glove box to keep the air moving.

Inoculating

The inoculation of your agar will require a sterilized environment (still air box/flow hood) to be successful. There is one exception to this rule, which is when you are transferring liquid culture into jars with airport lids, because the agar will not be exposed to any air at all during this process.

From liquid culture

A syringe will be used to transfer the liquid culture to the agar plate. Your liquid culture will either already be in a syringe, or it will be in a jar that you will need to transfer to a syringe before you can begin using it. Regardless of the method you use, you’ll need to stir your liquid culture to break it up so that each drop of liquid has a little amount of mycelium. Squirt a few drops of the liquid culture over the agar using the syringe; you won’t need much of it. If your agar culture is equipped with airport lids, you may do this procedure in a non-sterile setting.

From agar

Make sure you have a sterile syringe or a dental pick on hand before you begin. Cut a tiny wedge from the colonised agar and transfer it to new agar as rapidly as possible before sealing up the container. For petri dishes, it is a good idea to either place them in zip-lock bags or wrap parafilm around them to prevent them from drying out and becoming exposed to pollutants throughout the experiment.

From a mushroom

Agar culture creation by cloning a fresh mushroom is a basic process that is made easier if the mushroom that you are cloning is large. Alternatively, dip your mushroom into a mild H202 (hydrogen peroxide) solution for a few seconds, or spritz it liberally with a combination of water and methylated spirits. With your gloves on, split the mushroom in two using your hands to reveal the tissue in the center of the mushroom’s interior. Use a scalpel or a dental pick to cut a very little piece of tissue from the center of this tissue and transfer it to your agar plate for later use.

It is preferable to obtain a piece of tissue that has not been exposed to air because this will considerably lower the likelihood of pollutants being transferred over. You should be able to notice clear evidence of development within a few days following the transfer.

From spores

To transfer spores to agar, you will need to utilize an inoculation loop, which may be found here. You can get these for a reasonable price on the internet, or you may create one yourself out of some wire. Scrape the inoculation loop carefully over the top of the spores, and then pull it in a S form onto the agar using the loop (this is called streaking). The most spores should be found at the beginning of the streak (at the top of the S), and the least amount should be found at the end. The mycelium should begin to develop in the shape of a S within a few days to a week after you’ve planted the seeds.

You may either move the agar directly to grain spawn (multi-spore culture) or clip off robust appearing sectors of mycelium to transfer to fresh agar plates after your spores have germinated.

In order to make your own strain as a home cultivator, it’s ideal to start with multi-spore spawn, fruit the culture, and then clone the finest-looking mushrooms from the spawn.

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