Organism Insulation

To get the pure breeding mycelium of button mushroom, must be done insulation from tissue or spore or get the subculture from pure breeding which available.

Substance and appliance which is needed to tissue insulation and multiply the pure breeding as follows:

1. Gel medium to sterile and oblique in bottle or in reaction tube.
2. Alcohol lamp, scalpel, scissor, forceps, needle with the handle which do not conduct heat.
3. Clorox solution 5 - 10 %
4. Box or lint 50 x 50 cm.

Mushroom isolation should be done in the isolation room or on isolation box that provided with UV lamp for sterilization. If those appliances are not available, isolation can be done on the ceramic table that pillowed with lint, the lint is wetted with Clorox 5% first. Isolation room or isolation box must be sprayed with clorox first before used or other antiseptic solution as alcohol 40%.

Before isolation, hand must be sterilized with alcohol 40%. If using UV lamp for sterilize, light on the lamp one hour and turn off 0.5 hour before use for avoid UV effect to our body. On mushroom isolation can do for may purpose like tissue culture isolation, mono culture or multi spore, or multiply from pure culture from mycelium bank.

Multiply the Pure Breeding
After pure breeding of insulation have tested, pure breeding is ready to multiply in reaction tubes with PDA medium. This pure cultivated can directly use or keep. This technique usually is very difficult because it's always contaminated with environment. Other mushroom spore always there, you can't see this. Bacteria and virus also always available on air, so even we open just a bit, outside air will contaminated the media.