Procedures

Petri Dish To Petri Dish or Liquid Culture

All work should be conducted on a sterile surface in front of a HEPA-filter sterile air source. Read Stamets "The Mushroom Cultivator" or "Growing Gourmet and Medicinal Mushrooms" for details on sterile culture technique.

o) Prepare potato water or malt extract agar petri dishes or liquid cultures using our recipes or purchase petri dishes to inoculate

i) Sterilize all reused tools in autoclave(pressure cooker) when possible. Wash hands and arms before starting. Use a surgical mask and gloves while working. Rub hands down with 70 % IPA every 5 minutes or so. Remember not to use lighter while you still have IPA on hands!

ii) Clean work area with 70 % IPA. Wait until IPA fumes dissipate before using ignition source (torch, alcohol lamp, or bacti-cinerator)

iii) Arrange source culture and culture medium to be inoculated on working area; partially open any lids/remove parafilm so that rest of work can proceed swiftly. Only partially open the cultures you plan to use with one source culture at a time (2-5 petri dishes and/or 1-3 liquid cultures per petri dish with one sterilization of scalpel)

iv) Sterilize scalpel/inoculating loop with heat source. Keep IPA away! Invisible IPA fumes may explode/ignite! Be sure to have read the MSDS for IPA and follow safety precautions.

v) Cool scalpel in one of the containers of fresh sterile medium to be inoculated. Cut a tiny triangle from the agar of the colonized petri dish. Ascetically transfer to fresh petri dish. With graceful fast technique it is possible to do a few transfers before re-sterilizing the scalpel. As soon as you set the scalpel down to sterilize again seal any completed containers. To prevent contamination over longer term wrap with a thin strip of parafilm.

vi) You can repeat iv-v several times to get more transfers, but the more the source petri dish is exposed the higher the chance of introducing contamination.

vii) Incubate at correct temperature for species

To Grain Spawn

We recommend transferring petri dish cultures to liquid culture and using the liquid culture to inoculate grain bags, or using our culture suspensions to inoculate grain bags. This "double wedge" technique is often adequate for fast growing species like pink oyster and wood oyster (P. ostreatus) species

o) Prepare sterile grain 5-6lb 3T type filter patch bags using our recipes or purchase sterile grain in 3T filter patch bags to inoculate

ii) Clean work area with 70 % IPA. Wait until IPA fumes dissipate before using ignition source (torch, alcohol lamp, or bacti-cinerator)

iii) Arrange source culture and grain bags to be inoculated on working area; partially open any lids/remove parafilm/unroll and open bags so that rest of work can proceed swiftly. Only partially open the cultures you plan to use with one source culture at a time ( 1-3 grain bags per petri dish)

iv) Sterilize scalpel with heat source. Keep IPA away! Invisible IPA fumes may explode/ignite! Be sure to have read the MSDS for IPA and follow safety precautions.

v) Cool scalpel by contacting with sterile grain to be inoculated. Cut a large (2.5 cm) triangle from the agar of the colonized petri dish. Ascetically transfer to grain bag. Then transfer another triangle to same bag. With graceful fast technique it is possible to do a few transfers before re-sterilizing the scalpel. You can inoculate 1-3 bags with 2 wedges each before resting to resterilize the scalpel after sealing the bags with an impulse sealer. Label your sealed bags.

vi) Repeat step (v) until all your grain bags are inoculated. Shake the sealed bags to spread the wedges and distribute moisture evenly. Incubate in a moderately humid room at the correct temperature for the species

Liquid Culture To Grain Spawn

All work should be conducted on a sterile surface in front of a HEPA-filter sterile air source. Read Stamets"The Mushroom Cultivator" or "Growing Gourmet and Medicinal Mushrooms for details on sterile culture technique.

o) Prepare sterile grain 5-6lb 3T type filter patch bags using our recipes or purchase sterile grain in 3T filter patch bags to inoculate . Prepare liquid cultures from petri dishes using our instructionsand let them grow for 1-3 weeks until surface is totally covered in mycelium. For faster growth of grain spawn sanitize outside of lids by misting with 70 % IPA and shake liquid cultures 2-4 days before use.

ii) Clean work area with 70 % IPA.

iii) Sanitize tops of liquid cultures by misting with 70 % IPA

iv) Shake liquid culture quickly and vigorously.

v) Arrange source culture and grain bags to be inoculated on working area; partially open any lids/remove parafilm/unroll and open bags so that rest of work can proceed swiftly. Only partially open the cultures you plan to use with one source culture at a time (1-5 bags per ~200 ml liquid culture)

vi) Carefully pour equal portions of liquid culture into each grain bag you have opened. You can inoculate 1-5 bags with 200 ml of liquid culture before sealing the bags with an impulse sealer. Label your sealed bags.

vii) Repeat step (vi) until all your grain bags are inoculated. Shake the sealed bags to spread the liquid culture and distribute moisture evenly. Incubate in a moderately humid room at the correct temperature for the species

Culture Syringe To Petri Dish or Liquid Culture

All work should be conducted on a sterile surface in front of a HEPA-filter sterile air source. Read Stamets"The Mushroom Cultivator" or "Growing Gourmet and Medicinal Mushrooms" for details on sterile culture technique.

o) Prepare potato water or malt extract agar petri dishes or liquid cultures using our recipes or purchase petri dishes to inoculate

ii) Clean work area with 70 % IPA. Wait until IPA fumes dissipate before using ignition source (torch, alcohol lamp, or bacti-cinerator)

iii) Arrange source culture and culture medium to be inoculated on working area; partially open any lids/remove parafilm so that rest of work can proceed swiftly (if inoculating "over the top"). Only partially open the cultures you plan to use with one source culture at a time (3-10 petri dishes and/or 2-3 liquid cultures per culture syringe)

iv) Instead of squirting the syringe in "over the top" you can also use the needle provided and inoculate through an injection. In this case prep the area you will inject by rubbing with the IPA swab provided instead of loosening tops.

v) Squirt 0.5 ml to 3 ml per petri dish, or about 1-5 ml in each liquid culture. Close the new cultures immediately after using up syringe contents. To prevent contamination over longer term wrap with a thin strip of parafilm. If you injected through a bag seal the hole with 3M micropore tape or packing tape.

vi) Incubate new cultures at correct temperature for that species

To Grain Spawn

All work should be conducted on a sterile surface in front of a HEPA-filter sterile air source. Read Stamets"The Mushroom Cultivator" or "Growing Gourmet and Medicinal Mushrooms" for details on sterile culture technique.

This technique will work with most species.

For slow growing species (Maitake ect) it is best to make a larger liquid culture and use it to inoculate the grain spawn

o) Prepare sterile grain 5-6lb 3T type filter patch bags using our recipes or purchase sterile grain in 3T filter patch bags to inoculate

ii) Clean work area with 70 % IPA. Wait until IPA fumes dissipate before using ignition source (torch, alcohol lamp, or bacti-cinerator)

iv) Either squirt the scalpel into the top of the bag, or rub the side of bag with IPA swab and inject the syringe. Seal the top of the bag with an impulse sealer, and repair the injection site with micropore tape or packing tape. Label your bags.

v) Repeat step (iv) until all your grain bags are inoculated. Shake the sealed bags to spread the culture from syringe and distribute moisture evenly. Incubate in a moderately humid room at the correct temperature for the species

Grain Spawn To Sawdust Spawn or Sawdust/Bran Fruiting Mix

All work should be conducted on a sterile surface in front of a HEPA-filter sterile air source. Read Stamets"The Mushroom Cultivator" or "Growing Gourmet and Medicinal Mushrooms" for details on sterile culture technique.

It is best to grow a generation in sawdust and use those bags to inoculate the bran/sawdust bags. Mixing grain spawn into bran/sawdust mixture can cause excessive thermogenesis and overheating then contamination and death of mycelium. An incubation temperature 2-3 deg C below the recommended temperature for the species being used can help when inoculating grain into sawdust/bran.

o) Prepare sterile sawdust 5-6 lb A type filter patch bags using our recipes or purchase sterile sawdust in filter patch bags to inoculate

ii) Clean work area with 70 % IPA. Wait until IPA fumes dissipate before using ignition source (torch, alcohol lamp, or bacti-cinerator)

iii) Thoroughly break the grain spawn bag(s) down into individual grains by massaging through the plastic without puncturing. Sanitize the outside of the bag and a pair of scissors by misting with 70 % IPA.

iv) Arrange source culture and sawdust bags to be inoculated on working area; partially open any lids/remove parafilm/unroll and open bags so that rest of work can proceed swiftly. Only partially open the cultures you plan to use with one source culture at a time (5-10 sawdust bag per grain bag)

v) Cut a 2-5 inch corner off the grain bag with sanitized scissors. Pour equal amounts of grain into the sawdust bags. Once grain is used up seal bags immediately with impulse sealer. Label your bags.

vi) Repeat step (v) until all your sawdust bags are inoculated. Shake the sealed bags to spread the grain and distribute moisture evenly. Incubate in a moderately humid room at the correct temperature for the species

Sawdust Spawn To Sawdust/Bran Fruiting Mix

All work should be conducted on a sterile surface in front of a HEPA-filter sterile air source. Read Stamets"The Mushroom Cultivator" or "Growing Gourmet and Medicinal Mushrooms" for details on sterile culture technique.

This is the most reliable method to produce sawdust/bran fruiting kits.

o) Prepare sterile sawdust/bran 5-6 lb A type filter patch bags using our recipes or purchase sterile sawdust/bran in filter patch bags to inoculate.

ii) Clean work area with 70 % IPA. Wait until IPA fumes dissipate before using ignition source (torch, alcohol lamp, or bacti-cinerator)

iii) Thoroughly break the sawdust spawn bag(s) down into small particles by massaging through the plastic without puncturing. Sanitize the outside of the bag and a pair of scissors by misting with 70 % IPA.

v) Cut a 2-5 inch corner off the sawdust bag with sanitized scissors. Pour equal amounts of sawdust into the sawdust/bran bags. Once sawdust is used up seal bags immediately with impulse sealer. Label your bags.

vi) Repeat step (v) until all your sawdust/bran bags are inoculated. Shake the sealed bags to spread the sawdust and distribute moisture evenly. Incubate in a moderately humid room at the correct temperature for the species

To Logs (outdoors)

o) select hardwood logs which are not moldy and have fallen recently (last 3 months). Very fresh logs (less than one week since cutting) can also reject mushroom mycelium. logs should be 4-12 inches diameter, 2-5 feet long. Maple and Oak work well with most species. Most species require bark cover to form primordia, so to not use logs which have been stripped of bark.

i) drill the logs with ~2 inch deep holes every 3-5 inches in a diamond or grid pattern over the entire log, using a 12 mm drill bit.

ii) Open your sawdust spawn bag from the top. Use a spawn plunger tool . Push the plunger into the top of the spawn until the tool is filled, then use the plunger to inject into the holes you have drilled.

iii) Once the holes are filled melt low temperature soy wax and use a dabber to seal the spawn in the hole from attack by insects.

iv) When all the holes have been inoculated incubate the log outside just off the ground (on a skid), wetting occasional for the first half year. This can be done in a basement or garage in the winter. when you can see the mycelium reach the ends of the log it can be planted as totem (lion's mane, oyster), stacked and soaked (shiitake), planted into pots (reishi), or fully/partially buried (pioppino, maitake, kuritake) and left to fully colonize. Soaking the logs in the fall/spring can help bring on flushes of mushrooms. Once colonized the logs should flush for multiple years.

To mushroom beds (outdoors)

o) acquire fresh hardwood chips. A range of different sizes mixed together works better than uniformly sized chips. Maple and Oak work well with most species. The chips should be between 1 week to 3 months old, and not yet colonized by other molds.

i) select a shady, moist area.

ii) clear other fungi/weeds and cover the area with a layer of cardboard.

iii) lay out chips to a depth of 1-4 inches

iv) break up sawdust spawn and spread atop chips in a thin layer. Use 1 bag sawdust spawn per 1-8 square meters area. More spawn will lead to faster colonization and a greater success rate, but the same yield of mushroom fruit.

v) you can repeat steps iii-iv one or two more times to make a thicker mushroom bed that will produce for more years.

vi) cover last spawn layer with 1-3 inches of chips and wet the bed down to saturation. High chlorine content in water may kill your mushroom mycelium.

vii) you can put a layer of chicken wire to protect the bed from digging animals. Atop this put a layer of cardboard and/or FRESH straw to help the bed maintain moisture. Since growth of grass can help the growth of mushrooms, plant grass seed atop the bed at a low density (use only 10 % the recommended amount of grass seed for the area).

viii) you can mostly leave the bed to grow. during drought add water to prevent mycelium dying from desiccation. Soaking the bed in the spring/fall can elicit flushes.

Fruiting Kit:

Refer to species specific instructions on species page.

Finished fruiting kits can be composted or inoculated into logs or outdoor beds in the same manner as sawdust spawn