The following is a tried and tested method for maximizing the use of your myco-cultures while keeping contaminants to a minimum, and maintaining ease of innoculation.
Once you have purchased you selected species/variety in myco-culture form, the next issue is how to use this contamination free, isolated mycelium to begin innoculation of your selected sterilized/pasturised substrate.
Dutchie's BABY JAR METHOD is a easy way to produce a mycelium rich water solution in a syringe, used to innoculate suitable substrates. remember "STERILITY IS THE KEY!" any other sterile methods such as spraying the work area with disenfectant such as glen20 or lysol is recommended.
What you need:
1X Small cleaned and sterile baby jar with suitable re-closeable resealable lid.(larger 250ml jars may be used also)
1X 10ml sterile syringe with sterile long sharp/needle
10-20ml sterile water
1X Myco-culture
1X sterile scaple or hobby knife
suitable steriziling procedure(pressure cooker or double boiler)
Sterile/alcohol swabs
Heat resistant tape-plumbers tape(duct tape)
optional: Honey.
What to do:
Take the lid of the baby food jar an punch a small hole in the centre, around 2-3mm dia. Clean hole with alcohol swab and place a peice of tape over the hole, rub down securely.
Ensure the inside of the baby food jar is very very clean, wipping jar with swabs after it is cleaned. Add sterile water. Addition of growth suppliments can be added now, eg. Malt extract, honey etc..(option: add 1 part honey per 50 parts water)Place lid on jar loosely.
Place jar with lid on in top section of double boiler, and allow to heat up at boiling temp for around 20- 25 minutes to ensure all contaminants are destroyed (pressure cooker is optimal here, 15minfull pressure). Carefully remove jar and secure lid tightly while hot, ensuring tape is secure over the punched hole. Most good tapes will hold the needed pressure without splitting or coming undone. Use a better brand of tape to prevent problems.
Allow the jar/water to cool to room temp.
Take myco-culture and carfully remove-cut along the tape to free the lid, Do not open yet! If possible, work in a glovebox or over an indirect heat source, such as an open oven or stove, the hot air rises and prevents contaminants 'falling in'.It helps marginally , but it is better than not using this method. Be careful with disenfectants and heat sources, these can be flamable. Do not allow the heat to directly warm the culture, we only want the warm air to rise here, not to cook the culture.
The next step should be done as quickly and efficiently as possible. Please read over carefully before attempting this procedure...
Take sterile scaple or hobby knife in one hand, with the jar lid slightly loose and myco-culture ready to open. Have all items in close proximityof eachother, to minimize 'travel' time.Open the culture only slightly, cut a peice of mycelium rich agar from the myco-culture plate around 10-20mm square or so and 'stab' it with the knife to pick it up and close the plate. Quickly while over the warm air, transfer the cut section to the baby jar(now cooled to room temp)and quickly close the jar. Now you have transfered a peice of mycelium rich agar to the jar. Re-tape the myco-culture with plumbing tape or more micro-pore tape.
Shake the jar vigourously for around 3-5 minutes to ensure even distribution of mycelium.
Option 1: Draw up some mycelium water and while still holding a swab over the needle still in the hole, squit the water over the face of the agar, this will blast off more mycelium. Repeat a few times to distrubute the mycelium, shake again. Now you have an even, mycelium rich water you can use for innoculation. Retape if not all being used.
Option 2: Keep the innoculated jar in a suitable incubator or at constant room temp. Maximum mycelium growth will occur over the next 5-10 days with the addition of honey to the water. Shake vigourously for a few seconds each day to airate the water/honey/mycelial slurry mix. The extra few days growth insure large amounts of fine mycelial growth suspended in a rich medium. Once enough mycelium has formed (some cultures will grow for 4-5days some 20+) inject 1-2ml/250ml of hyrogen peroxide into the jarsand shake to eliminate any unwanted contaminats. Allow the slurry to recover for a few hours to a day and inocculate suitable substrates. Maximum colonisation time and nill conatminats has been acheived by using this method.
Use this great mycelium rich water to innoculate your favourtie substrates using around 1/2-2ml per innoc.site. Occasionaly if you add too much mycelium water you can change the composition of the substrate, so be careful.
