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How to Use Your Master Agar Slant
Getting Ready to Use Your Master Slant
Master Slants are meant for long term storage. The culture we use in our master slants is the lowest possible transfer value possible. This means we used youngest, healthiest mycelium to colonize the master slant. The reason for this is so you can go back to your master slant at a much later date and expand it to refresh the culture you use to inoculate with.
After about 15 - 20 transfers the mycelium will begin to weaken and may even eventually reach senescence, This will render the culture unusable at some point. The remedy to this problem is to always have a back up master slant that is colonized with a young and healthy culture specimen.
To start we will need to gather up some basic supplies.
- Fully colonized master slant.
- Some rubbing alcohol or sanitizer to clean with.
- A blank agar plate to transfer the mycelium to.
- A nice scalpel, one with a long skinny handle helps.
- A propane torch or spirit lamp to flame sterilize your scalpel.
- Some para film or saran wrap to reseal the master slant when finished.
Get Your Slant Ready
Take the parafilm off the master slant.
Wipe the master slant with sanitizer or alcohol.
Wipe the master slant with sanitizer or alcohol.
Your fully colonized master slant will arrive with the cap wrapped with some parafilm. You will need to remove it. Find the end and unroll it.
Wipe the master slant with sanitizer or alcohol.
Wipe the master slant with sanitizer or alcohol.
Wipe the master slant with sanitizer or alcohol.
Now you will fully wipe down the master slant with rubbing alcohol or the equivalent. Most sanitizers will work. We need the test tube clean.
Sterilize the Scalpel Blade and Open the Master Slant
Using a flame, sterilize the scalpel blade.
Using a flame, sterilize the scalpel blade.
Using a flame, sterilize the scalpel blade.
Flame sterilize using a propane torch or a spirit lamp with denatured alcohol. Do not use a lighter or a candle; they will leave carbon residue.
Unscrew the lid on the master slant.
Using a flame, sterilize the scalpel blade.
Using a flame, sterilize the scalpel blade.
Set the scalpel down to cool and unscrew the cap on the master slant. Make sure to keep the opening facing the flow hood filter.
Insert the Scalpel and Transfer the Mycelium
Insert the scalpel into the master slant.
Insert the scalpel into the master slant.
Insert the scalpel into the master slant.
With your flame-sterilized scalpel, carefully insert the blade first. Make a small scooping motion cut to get some mycelium on the scalpel.
Pull out a piece of mycelium.
Insert the scalpel into the master slant.
Insert the scalpel into the master slant.
Now that your mycelium has been exposed, keep it in the sterile air flow. Transfer the mycelium to a new agar plate for expansion.
Attach the Lid and Finish Up
Reattach the lid to the master slant. Wrap the lid in parafilm.
After you have transferred your mycelium, reattach the lid on the master slant. Screw the lid down tight, then turn the lid back 1/8th of a turn, just so it is the ever so slightly loose, but not loose enough to wiggle around when touched. Wrap the lid and test tube with para film.
Store your master slant in cool room temps for up to 1 year. It is best practice to make a new master slant annually. Here at Mushroom Cultures we make 2 slants and store them in different places.
Make sure to clean up your work area and put all of your tools away. A mycology lab is not the place to leave a mess and have disorganized. A messy lab is a dangerous lab. Keep it clean!