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PsilocyBen17
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Spore print on Agar
#19236557 - 12/06/13 01:22 PM (10 years, 1 month ago) |
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Hey guys,
I've done some reading on Agar and recently recieved 5 plates from The Mushroom Kingdom. I'm currently growing B+ and don't have any MS left. Instead of making a spore print and making that into a spore syringe (extra steps, meaning extra chance of contams) I was wondering if I could drop a fresh, sterile cap directly on the agar and wait for the spores to drop. Any advice if I choose to do this? I've also read that I could place a cap directly into a jar of substrate and achieve the same result.
Thanks PsilocyBen17
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shopdropper
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the cap, no matter how clean you think it is, is NOT sterile. just make a nice clean print on foil or paper and scrap those spores onto the agar in a still air box. save the other plates incase you have to out run some contams.
good luck
-------------------- DON'T TRY THIS AT HOME: no guarantees can be made about the accuracy of the information herein. The information dicussed in these posts is purely hypothetical, and for intelectual purposes only. Any similarity between internet chat and real life is pure coincidence.
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PsilocyBen17
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Quote:
shopdropper said: the cap, no matter how clean you think it is, is NOT sterile. just make a nice clean print on foil or paper and scrap those spores onto the agar in a still air box. save the other plates incase you have to out run some contams.
good luck
Thanks shop. I never thought of a print of foil. That will save me some steps.
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fatchillin
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Or you could take a fresh fruit you have, and clone a piece of the inside of the stem onto agar. Then you can clone a cluster or big fruit you'd like to have. MS is ok, but isolating is where it's at :P have fun with it
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Amanita virosa
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Registered: 12/04/11
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Quote:
fatchillin said: Or you could take a fresh fruit you have, and clone a piece of the inside of the stem onto agar. Then you can clone a cluster or big fruit you'd like to have. MS is ok, but isolating is where it's at :P have fun with it 
Exactly. Find your best fruit and clone it. Remenber to wipe it down with a bit of Iso and then tear it open with gloved hands in an sab. Never cut it open. Cutting drags surface bacteria into the inner flesh. Then use a flamed and very sharp scalpel to take a tiny piece of upper stem tissue and drop it on agar. Waa laa!
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MudaFuka
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I would definitely take a couple of clones but it allso never hurts to have a few prints around for when your mycelium gets to old and worn out.
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oONoodlesOo
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Quote:
PsilocyBen17 said:
Quote:
shopdropper said: the cap, no matter how clean you think it is, is NOT sterile. just make a nice clean print on foil or paper and scrap those spores onto the agar in a still air box. save the other plates incase you have to out run some contams.
good luck
Thanks shop. I never thought of a print of foil. That will save me some steps.
(<.<") here. http://www.shroomery.org/11342/Spore-Prints-and-Syringes
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themckid
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IMO one of the best ways to do agar work is from pins!
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PsilocyBen17
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Re: Spore print on Agar [Re: themckid]
#19403499 - 01/11/14 12:02 PM (10 years, 19 days ago) |
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So you mean do a biopsy from a pin? New to agar here.
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Amanita virosa
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yes he means using a mushroom primordia rather than a mature mushroom for the tissue culture. They are often cleaner. Just remember to tear not cut the fruit you are going to clone
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PsilocyBen17
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How would i tear a pin open? Its so tiny. Sorry for the newb questions just dont want to waste expensive agar plates
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Amanita virosa
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well... cut it at the base and tear it in two by grabbing either side of the cut edge. let it get big enough to handle somewhere between pin and fruit
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Aleon
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Quote:
PsilocyBen17 said: How would i tear a pin open? Its so tiny. Sorry for the newb questions just dont want to waste expensive agar plates
Using your fingers.
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Kizzle
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Re: Spore print on Agar [Re: Aleon]
#19410400 - 01/12/14 10:12 PM (10 years, 18 days ago) |
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You don't need to tear open the pin when you're cloning a tiny pin like that. You can just place it on the agar and make another transfer shortly after from the outer edge of the fresh mycelium growing across the agar. That should be free of contaminants and result in a clean culture.
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Aleon
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Re: Spore print on Agar [Re: Kizzle]
#19411359 - 01/13/14 06:15 AM (10 years, 18 days ago) |
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Quote:
Kizzle said: You don't need to tear open the pin when you're cloning a tiny pin like that. You can just place it on the agar and make another transfer shortly after from the outer edge of the fresh mycelium growing across the agar. That should be free of contaminants and result in a clean culture.
That also works; i have had to do it for cloning thin or woody species like turkey tail. I just placed a piece of the mushrooms on agar.
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supernewfie
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Re: Spore print on Agar [Re: Kizzle]
#19428907 - 01/16/14 04:06 PM (10 years, 14 days ago) |
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supernewfie
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Ive used an old syringe for all my agar/pin cuttings.. flame sterilize the needle and cut out the agar slice, or if it is a pin i plucked the pin up by placing the top of the needle under the base and wedge it up. I pierced the base of the pin to transfer it into the agar, in my case pancake mix.
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PsilocyBen17
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Woa woa woah...you made agar from pancake mix?!
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supernewfie
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Yes never had agar. Cronicr has a good thread about using BRF, me not having agar or brf, and I already knew the pancake mix worked since I made a cake and it colonized quickly, I used it and as you can see, its working.

This pancake mix is whole grain and organic ... it aint no aunt jamimas lol
Edited by supernewfie (01/17/14 08:41 AM)
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PsilocyBen17
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So the Pancake mix is your substrate...not your agar...
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PsilocyBen17
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Im confused by what your saying. Can you post the article?
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supernewfie
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Im using it to grow out mycelium from pins from agar plates, because I havent got agar. Cronicr has a thread where he used BRF.. it worked for him .. I never had brf either but I do now so il try some of that and see what happens. Il transfer the mycelium to a grain jar or something when it grows out
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supernewfie
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supernewfie
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stevo

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Re: Spore print on Agar *DELETED* [Re: supernewfie]
#19462301 - 01/23/14 12:17 PM (10 years, 7 days ago) |
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Post deleted by stevo
Reason for deletion: .
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Rabbit88



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Re: Spore print on Agar [Re: stevo]
#19504576 - 02/01/14 03:03 AM (9 years, 11 months ago) |
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Thats crazy ive never once even thought of using pancake batter as agar type substitute I guess u must adapt and use the resources around you
have you ever not added enough water and made bread in a mason jar while pressure cookin it?
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supernewfie
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Re: Spore print on Agar [Re: Rabbit88]
#19505581 - 02/01/14 10:32 AM (9 years, 11 months ago) |
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I put so much water that it never hardened its still pretty much liquid. Also the pancake mix is organic and uses a number of grains thats used in growing mushrooms.
Also I dont have a pressure cooker, I use a pot and lid to sterilize
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PsilocyBen17
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I wasn't able to find the info in the thread you've posted 
Do you just take organic pancake mix, add water, sterilize it in jars, and put some myc or a pin in there after it cools?
Do you use anything as a filter?
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supernewfie
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The thread im talking about is called my 50 dollar adventure, cronicr posted. And yeah water, pancake mix stir it up good make sure its not too thick caise it will turn to pancake when sterilizing if theres not enough water in it.
I dont use a filter.
BRF was used originally by cronicr
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supernewfie
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The thread I posted, page 1, cronicr explains how he does it. I suggest you follow this technique and use brf. I done this without recording weight of mix and amount of water so I wouldnt be able to tell you what to mix but in his post he gives a good explanation of how to set this kind of experiment up.
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PsilocyBen17
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Ah I see. He just uses a BRF/water mix as a substitute for agar. How long would you recommend steam sterilizing for? Thanks for your help
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supernewfie
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I sterilized for about 45 minutes to an hour. I checked it a few times to make sure it never solidified and added boiled water from the kettle when it did. I was just moreover fucking with it, experimenting... I used plastic microwavable containers.
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