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nn-IlliniSpiralDMT
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Agar Issues
#18926258 - 10/03/13 02:57 PM (10 years, 3 months ago) |
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How long on average does it take for an isolate to take on a new type of agar? I place an agar piece on a new agar plate on 9-29 and it has not even begun to grow? The plates look clean, but some have condensation. I just cant seem to avoid these problems, how the fuck do I avoid this? My agar transfer is just growing on itself and not the plate. WHY?
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nn-IlliniSpiralDMT
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EDIT- I think I added too much agar to my mix around 15 grams for 650 ml of water, 10 grams dog food, 2 grams karo, 2 grams BRF. Are these ratios okay, my mycelium is unusually hard and rubbery, I tossed one of my plates and felt it.
Any idea
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llama_police
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That's a pretty complicated agar recipe. Why not just stick with a basic malt extract agar and use 4g of agar and ME per 100ml?
And to answer your initial question, you should have seen growth by now, yes. Only takes a few days if all your ducks are lined up.
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RogerRabbit
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Quote:
nn-IlliniSpiralDMT said: EDIT- I think I added too much agar to my mix around 15 grams for 650 ml of water, 10 grams dog food, 2 grams karo, 2 grams BRF. Are these ratios okay, my mycelium is unusually hard and rubbery, I tossed one of my plates and felt it.
Any idea
That's far too nutritious. Leave out the karo and brf next time. RR
-------------------- Download Let's Grow Mushrooms semper in excretia sumus solim profundum variat "I've never had a failed experiment. I've only discovered 10,000 methods which do not work." Thomas Edison
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nn-IlliniSpiralDMT
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Quote:
RogerRabbit said:
Quote:
nn-IlliniSpiralDMT said: EDIT- I think I added too much agar to my mix around 15 grams for 650 ml of water, 10 grams dog food, 2 grams karo, 2 grams BRF. Are these ratios okay, my mycelium is unusually hard and rubbery, I tossed one of my plates and felt it.
Any idea
That's far too nutritious. Leave out the karo and brf next time. RR
Wait LOL are serious!?
So should I have just used say the dog food? And could I possibly just dilute my flask by adding 250ml and PCing again? Im really screwed here, Iv'e been transferring this for 4 months and finally Iv'e gotten fruits to produce around 2 grams dry per agar dish and they seem stellar. Anyway I could just grab the mycelium off of the other plate that's growing and sand-which it between another new dish and fresh agar? Thanks again RR
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FrankHorrigan
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Quote:
nn-IlliniSpiralDMT said: Iv'e been transferring this for 4 months and finally Iv'e gotten fruits to produce around 2 grams dry per agar dish and they seem stellar
This makes no sense. What are you saying?
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mushrume man
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All i can guess is fruiting from agar. this is why OP uses so much nutrients?
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forrest



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than you would still have to have quite a lot of agar to have enough moisture for fruits of 20 grams. do you have pics OP?
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Stromrider
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Re: Agar Issues [Re: forrest]
#18934808 - 10/05/13 05:22 AM (10 years, 3 months ago) |
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Fruiting from agar? That doesn't sound like a very efficient method
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nn-IlliniSpiralDMT
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Quote:
FrankHorrigan said:
Quote:
nn-IlliniSpiralDMT said: Iv'e been transferring this for 4 months and finally Iv'e gotten fruits to produce around 2 grams dry per agar dish and they seem stellar
This makes no sense. What are you saying?
Let me clarify for you FRANK Awhile back I found an old spore print from a very old grow found in a Simpsons Christmas special DVD. I then used those spores to knock up a whole sleeve of petris in open air with a Qtip, don't give me shit for using Qtips, I use those in place of inoculating loops and they are completely sterile, I grab them from the hospital I work at. So anyway 5 of those in that sleeve ended up being contam free and from there I did a couple of transfers and ended up with 3 dishes that I liked. All three of those dishes were left in my storage container and fruited. I don't want to fruit them in agar dishes(that was not my objective) but they yielded 2 grams dry give or take per dish. I grabbed one of the fruits and did a tissue transfer from the inside stripe. I gave those dried specimens to a friend and he was very pleased with how 1gram of those fruits reacted with him.
To simplify I have been doing agar transfers for about two years and all the dishes I have in storage are okay, but none and I mean none come close to how this one particular plate has reacted with me and my friend. I mean straight  and never even indulge in such activities anymore. Basically I want to save this specimen but I cant because I used this fucking agar recipe and got fucked over. I never even knew that agar could be too NUTRITIOUS, so now Im left with 3 plates that is just growing on the transferred piece of agar.
SO PLEASE HELP and give advice.
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nn-IlliniSpiralDMT
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I wasn't purposely trying to fruit on agar, but from my experience its best to let your agar fruit by itself and see if it can even fruit before doing A2G and coming up with a tub full of shit
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nn-IlliniSpiralDMT
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Re: Agar Issues [Re: forrest]
#18935676 - 10/05/13 11:17 AM (10 years, 3 months ago) |
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Sadly I dont, but I think I might have one with a cluster pic, I'll look. To be honest I have no idea how to upload pics on here and feel sketchy doing so... but could I just upload them from my phone?? I got great pics of plenty of strains from cubes to shiitake to just random specimens found in the woods of Wisconsin and Illinois if your interested...
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SpitballJedi
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Quote:
nn-IlliniSpiralDMT said: I wasn't purposely trying to fruit on agar, but from my experience its best to let your agar fruit by itself and see if it can even fruit before doing A2G and coming up with a tub full of shit
My understanding is, and I could be wrong, if you fruit on agar then that culture is in fruiting mode and does not respond well when switching back to colonization mode.
If this is true, then you could avoid the issue by just fruiting a transfer from a master. Once you see that it fruits and is potent, then you can just go back to the master.
However, just because it does not fruit on agar does not mean it won't fruit on a properly prepared substrate.
The reason I say I could be wrong about my initial premise is because it seems to me that a clone from a fruit would also be in fruiting mode and a fruit is basically mycelium anyway. 
I can see your agar fruits being small and potent. I typically pick my fruits smaller than most people probably do. My friends and I have done some experimenting and have agreed that I should pick them before the veil breaks or get too big.
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Rauhfasertapete
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seems the mushroom doesn´t like substrate. I had something like that before with an excessive sugar content. did you use exactly the same recipe for your other transfers before?
15gagar/650ml is much, but some people use even more. agar actually doesn´t do any harm.
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nn-IlliniSpiralDMT
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Quote:
SpitballJedi said:
Quote:
nn-IlliniSpiralDMT said: I wasn't purposely trying to fruit on agar, but from my experience its best to let your agar fruit by itself and see if it can even fruit before doing A2G and coming up with a tub full of shit
My understanding is, and I could be wrong, if you fruit on agar then that culture is in fruiting mode and does not respond well when switching back to colonization mode.
If this is true, then you could avoid the issue by just fruiting a transfer from a master. Once you see that it fruits and is potent, then you can just go back to the master.
However, just because it does not fruit on agar does not mean it won't fruit on a properly prepared substrate.
The reason I say I could be wrong about my initial premise is because it seems to me that a clone from a fruit would also be in fruiting mode and a fruit is basically mycelium anyway. 
I can see your agar fruits being small and potent. I typically pick my fruits smaller than most people probably do. My friends and I have done some experimenting and have agreed that I should pick them before the veil breaks or get too big.
On the matter of when to harvest fruits I say it depends on what you find preferable, I agree, picking early is a better ratio for potency of cubensis. It isn't necessarily that the mushrooms are more potent, its more so along the lines of water dilution within that mushroom. If that makes sense. In most edible mushrooms its also better to pick before they mature, as some species seem to get rather rubbery and tough out on the outside.
Honestly I haven't heard of fruits on agar not producing a network of mycelium. Usually fruits are vigorous when placed around nutritious mediums but you could be right and the times I have done I've gotten lucky. Let me know if you can find anything on that matter.
And trust me, you can add too much agar. I think that's my culprit right now and RR pointed that out too. It feels like rubber, and the agar piece I used to inoculate the dish is growing rapidly. I might just make some more dishes, and transfer a couple of slices of the initial transfer, and maybe shoot up a couple of grain bags with some new Agaricus brunnescens MS I just whipped up. Im just going to do 2 1lb bags and case them with peat moss.
I'll get back to you and see if I can save this prized horse or if its going to end up in my bed like the Godfather HAHAHA
PEACE
IS
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nn-IlliniSpiralDMT
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No thats the problem, usually I buy premade mixes of PDY but I ran out and never ordered any. So I've been playing around in the kitchen trying to make new recipes. I think I'm going to stick to the OG recipe I have, let me know what you think.
250 grams of potatoes cut up and boiled in 500ml of water, boil for 10 minutes and strain. Removing the actual potatos and having a broth left. Then add 2 grams of karo and thats it- besides PCing. Is that too much karo in your opinion? I might use the dog food again but I dont like having too much sediment in my dishes.
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inski
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For 500ml I would use 10g agar, the rest of your recipe sounds alright although I use dextrose rather than karo.
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nn-IlliniSpiralDMT
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Re: Agar Issues [Re: inski]
#18938131 - 10/05/13 09:56 PM (10 years, 3 months ago) |
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Quote:
inski said: For 500ml I would use 10g agar, the rest of your recipe sounds alright although I use dextrose rather than karo.
Definitely will be adding 10grams Where do you usually get dextrose and what are the advantages, I thought karo is basically the same as dextrose which is just corn sugar. I was going to head to a local brew shop on Monday after class and grab some light malt extract.
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inski
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Registered: 02/28/06
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No, there is no problem with karo, I just haven't tried it before because I haven't seen it for sale anywhere.
I got my dextrose from a supermarket, sorry I can't help you, I'm in New Zealand.
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Rauhfasertapete
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Re: Agar Issues [Re: inski]
#18939149 - 10/06/13 07:34 AM (10 years, 3 months ago) |
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Here´s another potato agar recipe from a mushroom cultivation book, which always worked out for me:
300 grams potatoes 12 grams dextrose a tea spoon of beer yeast (I use 7 to 10 grams fresh moist baker´s yeast) 1,1 litre water (refill what has evaporated; I sometimes use some more water, but the original recipe is okay) 25 grams agar (I ussally use less then half)
dextrose is a monosaccharid, which is a bit easier to digest than the disaccharids saccharose or maltose, because the mushroom needs lots of enzymes to crack glycosidic bonds. Karo consists of dextrose and a few percent fructose (also a monosaccharid). Honey is also mainly monosaccharids.
And what os OG again?
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Rauhfasertapete
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By the way, in the same book there´s also a dog food agar recipe: 30g Pedigree-Pal dog food 25g agar 1,1 l water
also quite high in nutrients. I never tried it, but if such a recipe works, your own recipe´s nutrient content cant be so much out of range. could it be that there were some preservatives in your dog food?
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Edited by Rauhfasertapete (10/06/13 08:00 AM)
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Stromrider
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I have been using dog food agar lately and it has been working great
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nn-IlliniSpiralDMT
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Quote:
Rauhfasertapete said: Here´s another potato agar recipe from a mushroom cultivation book, which always worked out for me:
300 grams potatoes 12 grams dextrose a tea spoon of beer yeast (I use 7 to 10 grams fresh moist baker´s yeast) 1,1 litre water (refill what has evaporated; I sometimes use some more water, but the original recipe is okay) 25 grams agar (I ussally use less then half)
dextrose is a monosaccharid, which is a bit easier to digest than the disaccharids saccharose or maltose, because the mushroom needs lots of enzymes to crack glycosidic bonds. Karo consists of dextrose and a few percent fructose (also a monosaccharid). Honey is also mainly monosaccharids.
And what os OG again?
OG is slang for original gangster, like the first agar recipe I used was OG or my master jars are OG because they were the original. NO use for it really in mycology hahaha but I like to drop it every now anad then. I know the difference between dextrose(corn derived sugar), glucose(blood sugars), sucrose(table sugar), and fructose(a hexose sugar found in fruit usually). It is interesting to know why mycelium takes to certain medias though, thank you for that, so you're saying that mycelium cannot break down complex sugars but it's enzymes can break down monosaccharids.
Also that recipe isn't too rich in your experience? I've also read that yeast for baking is not as good as brewers yeast. I dont know why and I could be wrong but I remember reading something along the lines of that.
IS
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RogerRabbit
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Quote:
nn-IlliniSpiralDMT said: ll three of those dishes were left in my storage container and fruited. I don't want to fruit them in agar dishes(that was not my objective) but they yielded 2 grams dry give or take per dish.
That's almost the yield of a brf cake and nobody is going to get me to believe that could ever come from a petri dish. I've been doing this shit too long. . .  RR
-------------------- Download Let's Grow Mushrooms semper in excretia sumus solim profundum variat "I've never had a failed experiment. I've only discovered 10,000 methods which do not work." Thomas Edison
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nn-IlliniSpiralDMT
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Quote:
RogerRabbit said:
Quote:
nn-IlliniSpiralDMT said: ll three of those dishes were left in my storage container and fruited. I don't want to fruit them in agar dishes(that was not my objective) but they yielded 2 grams dry give or take per dish.
That's almost the yield of a brf cake and nobody is going to get me to believe that could ever come from a petri dish. I've been doing this shit too long. . .  RR

I respect what your saying, and to be honest I wouldn't believe it either if I were you but If it wasn't true I wouldn't be here trying to save this isolate You see that picture you have? my plates were like that except way thicker meatier specimens. Id say that plate you have is around .4grams dry. I didn't say exactly 20grams but around, If I remember correctly it was like 1.7dry and they were the best in quality. Listen I normally wouldn't give a crap about saving one little isolate, but IM TELLING YOU THIS WAS THE HOLY GRAIL! The isolate that everyone dreams about and I never even got to put it to grain and fruit. Which is why IM TELLING YOU I NEED TO SAVE IT. I've been doing this shit on my own for two years and I think it was just plain luck that I found this spore print to begin with and a lottery pick that I was able to isolate this sector. I've been to southern mexico and have seen Cubensis get as big as my fucking arm...
And c'mon RR you really only average 20 grams wet for a PF cake on an isolate? I don't believe that for one second, not with your experience bro...
But my plan is to whip up new plates and transfer the intial transfer to clean agar. But I think I have a slight bacteria problem growing around my plate. Probably from too much condensation from the plates cooling is my though, any success of transferring the initial transfer in your experience?
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
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Quote:
RogerRabbit said:

Sweet pinset
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FrankHorrigan
The Inquisition



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Quote:
nn-IlliniSpiralDMT said: Listen I normally wouldn't give a crap about saving one little isolate, but IM TELLING YOU THIS WAS THE HOLY GRAIL! The isolate that everyone dreams about and I never even got to put it to grain and fruit. Which is why IM TELLING YOU I NEED TO SAVE IT.

Sounds like you're bullshitting us.
I doubt your agar in the entire dish weighs 20g.
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Pastywhyte
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Hold on everyone I grew this offa 2 tablespoons of brf fert agar, fruited under a discoball and neon blue lights, harvested on a full moon and got 213 grams wet. It dried to 454 grams Just look at that canopy!

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Stromrider
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good stuff
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nn-IlliniSpiralDMT
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Quote:
Pastywhyte said: Hold on everyone I grew this offa 2 tablespoons of brf fert agar, fruited under a discoball and neon blue lights, harvested on a full moon and got 213 grams wet. It dried to 454 grams Just look at that canopy!


Yeah, everybody laugh
What keeps me smiling is I know what I have and you guys apparently do not
Sorry I got a homerun while you guys are still swinging, you know what's most disappointing though? I remember coming on here for the first time as a junior in high-school and trying to learn as much as I could and people would actually help. They might of poked fun, but they actually gave advice and often steered people in the right direction while giving hope that they one day could actually be a successful mushroom grower whether in medicinal or gourmet or psychedelic whichever you directly prefer. But now 4 years later as a junior at a University,everyone just acts like a know it all, like their shit doesn't stink why does everyone think they are GOD? What is the need for this internet macho aficionado that rules the forums like a tyrant beheading people for giving advice. I mean literally every time I give advice on something that works for me and has proven to give me good results I get bashed, I'm not on here wasting my time "making up shit" and "bullshitting" people for fun.
As for now, I really didn't think 17 grams on an agar dish was that rare and now I guess I need to just stay off this particular thread and get to work try to save this culture, get some pictures and prove you guys wrong..
Until then I'll stick to trying to fruit these button mushrooms on cased grain.
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FrankHorrigan
The Inquisition



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SpitballJedi
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Let me do a calculation just on water content.
20g agar + 20g LME + 1000g water = 1040 media.
This mix is ~96% water. 1mL water weighs 1g.
Petri dish: (poured at 3mm deep by 100mm wide) V=πr2h V=π502*3 V=23562mm3 V=23.5mL
My 23.5mL of media is 96% water. So therefore,
96% of 23.5mL of media= 22.5mL of water
In perfect conditions, you could theoretically get 17 wet grams on a dish because there is enough water.
However, in my experience, fruits stop growing long before the water supply dries up. It seems that low moisture content stops growth long before the media gets bone dry.
By my calculations, I believe your mushrooms would stop growing long before they get the size/quantity you claim because there would not be enough water.
Just out of curiosity, I just weighed a 100mm x ~2-3mm wedge and it weighed 27g.
This calculation doesn't even consider the fact of how little room there is for growth and the lack of FAE.
For now, I'm calling bullshit.
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