Going from a print

[Trip Dr]

Hey fellas, just want to clarify something. Im thinking of growing some gourmet edibles from a print. My biggest concern is that the print is not sterile. My thoughts were that the best way to go would be to make up a heap of sterile petris with agar and simply wipe some spores off the print and onto the agar using a sterile needle or something. If i did this with say 10-20 petris i was assuming that many would get contaminated, but hopefully i would get lucky and have one that did not, and could then use that one to start a new series of sterile cultures. Does this sound feasible, or is there an easier method when working from a non sterile print?

[occidentalis]

That is generally how it's done

You might be suprised how clean it actually is. You'll probably only need 3 or 4 plates to get one that's clean enough to take a subculture from.

Use a sterile cotton bud.

[gerbil]

You might also wish to incorporate some h202 into your agar just before pouring if that interests you, especially in the subculture stage; about 6ml of 3% per 500ml agar is around about it, can be more can be less.

[worowa]

H2O2 can kill spores, so be careful. I recently tried catching some king stropharia, and dropped spores onto peroxidated agar-about a week later, and most dishes had mould, but 1 appears clean with mycelium!

Good luck

[Hyphal]

Streak some agar plates, then as they germinate take a small piece of un-contammed myc from them and transfer this to another clean agar plate. Continue to make isolations like this until you grow only clean healthy myc.

[Zen Peddler]

Get glove box, get a scalpel, clean the glove box using alcohol wipes, spray something in it too kill airborn contaminants (the majority that exist), burn the scalpel head inside the glove box, use it to streak the spores across 20 or so sterile agar plates. Most will contaminate if its a wild print.

Transfer the ones that dont onto to h202 agar. If all contaminate, its worth trying with ones that have bacterial contams - that is the oily sluge - any that contaminate with molds are generally done.

[occidentalis]

spray something in it too kill airborn contaminants (the majority that exist), burn the scalpel head inside the glove box

good advice - although overkill IMO

but allow me to point out a rather obvious, but often overlooked, safety issue ;)

The 'something' you will be spraying in to kill airborne contaminants will probably be ethanol, metho, or some other hydrocarbon spray

and then you will be flame sterilising your scalpel.....

It happened to me and I had some bad burns on my arms...

[Zen Peddler]

LOL. Good point. After continual contaminations I went hardcore on their ass...

[ENtiTY]

Heya peeps,

Its been mentioned before that the procedures and protocols for ensuring sterility, found on/in various forums and publications are a tad extreme. I recommend that people new to mycology follow them to the letter until they get a practical understanding of the whole process and why each component of the process is important. When you are learning something new you should learn to do it the correct/ideal way from the start and then once you have an understanding, you can chop and change in a effort to improve the process according to your personal preferences. Each person and location is different so what works for one person at their place might not work for you.

That said here's what I find most user friendly.

Glen20 is a good option for de-nastifying your glove box. Also I find using a jet lighter to glow the scalpel/hobby knife blade outside the glove box followed by a quick swipe of an Iso soaked towel and placed inside the glove box while the blade is still hot is safe and sterile.

Good luck.