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DaCultivater
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Registered: 10/08/05
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spore print to agar... soak or no soak?
#5440224 - 03/25/06 02:26 AM (17 years, 11 months ago) |
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so i've read bolth and plan to do the sp to agar tomarrow...
so what do you guys think i should do, soak or not soak the spores first?
will the spore still germinate without soaking? ty for any advise
-cult
-------------------- as they said in some movie(i don't remember which one); a belief is a dangerous thing, people will kill over it; instead of believing you need to find your facts and then have an opinionated idea that you are willing to change in light of new evidence
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agar
old hand
Registered: 11/21/04
Posts: 9,056
Loc: Somewhere Else
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Re: spore print to agar... soak or no soak? [Re: DaCultivater]
#5440428 - 03/25/06 07:57 AM (17 years, 11 months ago) |
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No soak is needed. There is plenty of moisture, in the agar, to germinate spores.
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hyphae
born to grow
Registered: 12/13/02
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Re: spore print to agar... soak or no soak? [Re: agar]
#5440432 - 03/25/06 08:04 AM (17 years, 11 months ago) |
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Depending on how old the spores are they can take quite along time to rehydrate. Dehydrated spores will not germinate BTW. Most importantly if you are using relatively fresh spores (up to say 1yr.) you should be fine either way make sure you break the surface and deposit some of those spore "in" the agar this will be the first place germination will occur in most cases.
-------------------- Getting the most out of your casings!, A pinning strategy. Oyster Shell "Flour" $2 for 1lb. a hell of a deal Not what is overlay but rather what overlay is Gas Exchange vs. FAE "We all have priorities. I used a closet once setup a nice little lab trouble was all the shit that was in there ended up in the bedroom that pissed off the GF then I ended up dumping her as she was getting in the way of my sterile culture technique! Ya I got priorities too!!!"
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bluemawkishness
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Re: spore print to agar... soak or no soak? [Re: hyphae]
#5440466 - 03/25/06 08:36 AM (17 years, 11 months ago) |
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I never soaked them so far, no probs. I just rasped some of the print on the agar.
If you have a very week print only, soaking can be helpful, because once you have a solution of the print in steril water, you can inoculate much more petris than by just rasping.
Edited by bluemawkishness (03/25/06 08:41 AM)
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hyphae
born to grow
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Re: spore print to agar... soak or no soak? [Re: bluemawkishness]
#5440519 - 03/25/06 09:40 AM (17 years, 11 months ago) |
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Quote:
bluemawkishness said: I never soaked them so far, no probs. I just rasped some of the print on the agar.
If you have a very week print only, soaking can be helpful, because once you have a solution of the print in steril water, you can inoculate much more petris than by just rasping.
I am only referring to old dehydrated spores BTW. Also even weak prints have plenty of spores the water is purely a means of re-hydration at this point. As far as on the surface or breaking the surface this was a tip from pro that has made my agar work easier than ever, using it or not it's up to the individual but I have to highly recommend it as it works and is a fine touch added to ones agar repertoire. I totally agree diluting spores in water will extend them but in agar IMHO it is simply redundant.
-------------------- Getting the most out of your casings!, A pinning strategy. Oyster Shell "Flour" $2 for 1lb. a hell of a deal Not what is overlay but rather what overlay is Gas Exchange vs. FAE "We all have priorities. I used a closet once setup a nice little lab trouble was all the shit that was in there ended up in the bedroom that pissed off the GF then I ended up dumping her as she was getting in the way of my sterile culture technique! Ya I got priorities too!!!"
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impgl
CrimethINCspecial agent
Registered: 02/07/06
Posts: 2,462
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Re: spore print to agar... soak or no soak? [Re: hyphae]
#5440797 - 03/25/06 11:53 AM (17 years, 11 months ago) |
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so im unclear as to the germination process. you soak the old spores in distilled water, right? how much water and for how long? also, heres the part thats really a noodle scratcher for me: when in liquid form, how do i put it on the agar? thanks in advance!
-------------------- omg really?
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Liquidkick
H2O
Registered: 05/03/02
Posts: 2,635
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Re: spore print to agar... soak or no soak? [Re: impgl]
#5440895 - 03/25/06 12:28 PM (17 years, 11 months ago) |
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1 single drop of spore solution on the agar dish.
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hyphae
born to grow
Registered: 12/13/02
Posts: 6,228
Loc: the rain forests
Last seen: 12 years, 10 months
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Re: spore print to agar... soak or no soak? [Re: impgl]
#5440909 - 03/25/06 12:33 PM (17 years, 11 months ago) |
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First I would use a sterile syringes a sterilized shotglass and sterilized water. In a glovebox (still air) I would flame the needle or just wipe it down with alcohol and scrape some spores into the shotglass then inject the sterilized water from the syringe into the shotglass containing the spores and suck it back up let the syringe sit at room temperature for 1-2 weeks then in a sterile environment (GB) place a few drops on agar. How old are your spores BTW? Also a dirty print will leave you with many contams and isolating pure myc gets difficult.
-------------------- Getting the most out of your casings!, A pinning strategy. Oyster Shell "Flour" $2 for 1lb. a hell of a deal Not what is overlay but rather what overlay is Gas Exchange vs. FAE "We all have priorities. I used a closet once setup a nice little lab trouble was all the shit that was in there ended up in the bedroom that pissed off the GF then I ended up dumping her as she was getting in the way of my sterile culture technique! Ya I got priorities too!!!"
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impgl
CrimethINCspecial agent
Registered: 02/07/06
Posts: 2,462
Loc: california!
Last seen: 7 years, 5 months
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Re: spore print to agar... soak or no soak? [Re: hyphae]
#5441069 - 03/25/06 01:24 PM (17 years, 11 months ago) |
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thanks for the info. its a p cube (Tazmanian) that i bought in amsterdam. completely sealed and prepped in a sterile lab (or so boasts the labeling). i got it around june of 04. whattaya think??
-------------------- omg really?
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hyphae
born to grow
Registered: 12/13/02
Posts: 6,228
Loc: the rain forests
Last seen: 12 years, 10 months
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Re: spore print to agar... soak or no soak? [Re: impgl]
#5441194 - 03/25/06 02:14 PM (17 years, 11 months ago) |
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Well you'll have plenty of spores if ya want to make some syringes and try spores directly to agar. I will recommend making at least one syringe at this time as this is as clean as that print will ever be. I would use a 1/2pt. jar and sterilize the water in that, use distilled water for sure as any other can germinate the spores because of the nutes in them. Remember all you need is a fraction of those spores to germinate and your in business. Remember when making the hole in the lid place it near the edge so you can get every last drop when tilted!
-------------------- Getting the most out of your casings!, A pinning strategy. Oyster Shell "Flour" $2 for 1lb. a hell of a deal Not what is overlay but rather what overlay is Gas Exchange vs. FAE "We all have priorities. I used a closet once setup a nice little lab trouble was all the shit that was in there ended up in the bedroom that pissed off the GF then I ended up dumping her as she was getting in the way of my sterile culture technique! Ya I got priorities too!!!"
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DaCultivater
Stranger
Registered: 10/08/05
Posts: 289
Last seen: 15 years, 11 months
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Re: spore print to agar... soak or no soak? [Re: hyphae]
#5441327 - 03/25/06 03:07 PM (17 years, 11 months ago) |
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lol ty for th advise guys
very helpfull i shall do it TODAY!!!! no soak 20 new dishes hear i come
-------------------- as they said in some movie(i don't remember which one); a belief is a dangerous thing, people will kill over it; instead of believing you need to find your facts and then have an opinionated idea that you are willing to change in light of new evidence
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