|Posted on Thursday, November 01, 2001 - 03:05 pm:||
Culture source and cloning
Either acquire a spore syringe or get started from a fresh fruit body.
Spores can be injected into the whole grain recipe or the MMGG recipe.
It /seems/ that spores work better in the MMGG recipe, but more tests need to be made. After getting a strain started, you can farm it for a long time with the procedure below.
Take a healthy mushroom for cloning around the time that the veil breaks. You want one that is still growing vigorously, not an abhort. Place it in the transfer chamber. Avoid fruit with brown streaks on the stem. Use a sterilized Xacto knife and tweezers to extract a clean chunk of stem to add to sterile water in a mason jar.
This is done by holding the freshly picked shroom near the cap, and using the sterile knife to cut a ring around the stem about 2" from the bottom. Only cut a little way into the stem.
Use sterile tweezers to grab and peel the surface layer back toward the base of the stem until it resembles a peeled banana.
Then cut the stem just above the peels. Once you've removed the Al foil from the sterilized blade assembly and mason jar, You have a sterile table to work on for cutting off the stub.
Then slice off the clean chunk and let it fall into a mason jar (8 oz regular mouth) 2/3 full of sterilized water. The jar is prepared by covering with the metal blade assembly and seal from the blender and that is covered with Al foil and sterilized. The transfer is done in the glove box. For more details see the "Blender Usage" page.
It turns out that a regular mouth mason jar has the same size and thread as the plastic blender container part from an Osterizer blender (probably other brands as well), so after dropping in the piece of stem, remove the jar from the chamber, then screw on the base of the blender and proceed to blend for about 3 or 4 seconds at the fastest speed. Allow to stop, then repeat. Be sure to hold the jar while blending. When done, remove the blender base, put the jar (w/seal and blades) in the chamber, remove the blades and seal, then immediately cover with a new baggie (in the transfer chamber). Then draw syringes of the inoculant water by piercing the plastic bag. This procedure has proved to be very effective. For pictures of the details of the blender parts and mason jar assembly go to the Blender Usage page.
1) It is best to use a rubber band around the shroom water jar/baggie to keep out contaminants.
2) If one cuts the peeled chunk of stem almost all the way thru', then one can lift the blender parts just enough to stick the stem thru and finish severing the sterile chunk from the rest of the stem with the edge of the jar and the blender blade assy.
3) The shroom water has been stored for 1 week with no harm and it's gone bad in 2 days. I have no idea how long it can remain viable. It is MUCH better to make up the shroom water and use it immediately. The growth rates are incredible when the shroom water is fresh. One pint cakes (rice only) have been birthed 10 days after inoculation. 14 to 21 days is more common.
4) The knife and tweezers are sterilized in Al foil sheaths in the pressure cooker.
5) Eventually the cloned strain will lose stamina and stop producing and growing vigorously. It is a good idea to take a spore print and start a new culture every year or so (at least.) See the spore print procedure.
Main Page : The Glove Box : Cloning : The Substrate : Casing/Fruiting : Spore Collection : Blender Usage : Tips : Shooting Liquid Culture
(Message edited by admin on January 14, 2004)
Post Number: 10654
|Posted on Tuesday, January 13, 2004 - 02:21 am:||
Post Number: 343
Posted on Friday, January 09, 2004 - 11:55 pm:
As simple as it sounds the best method I have found, to get good tissue to clone, is to use gloves and tear the fungi down the middle lengthwise. Then using a flamed scalpel cut a small piece of tissue from the center of the cap. If you cut it rather than pull it apart, you will drag any nasties that are on the outside of the fungi into the tissue you want to clone. This was the best piece of advise anyone ever gave me. I have cloned a few store bought fungi like this.