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OfflineGregP
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Sterilizing with Agar
    #23768560 - 10/24/16 10:45 PM (7 years, 3 months ago)

I know I will receive some flak for not making my own but I just purchased some agar plates online.  I printed a bunch of spores and want to sterilize before I inoculate.  Any guidance would be extremely appreciated.


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Invisibledankington
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Re: Sterilizing with Agar [Re: GregP]
    #23768564 - 10/24/16 10:47 PM (7 years, 3 months ago)

sterilize what?
You take sterile swabs or an inoculation loop, and inside your SAB you swab/streak your plates. You sterilize a loop (if it's metal) with a flame, cool it in your agar, then touch the print and streak the plates.


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OfflineGregP
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Re: Sterilizing with Agar [Re: dankington]
    #23768575 - 10/24/16 10:53 PM (7 years, 3 months ago)

So is it better if I get a loop. Very new to this. Have been ordering pre made syringes but want to start doing it myself.

So after I do that collect the print from the agar plate and make a syringe w distilled water?


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Invisibledankington
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Re: Sterilizing with Agar [Re: GregP]
    #23768581 - 10/24/16 10:58 PM (7 years, 3 months ago)

you collect a print from a mushroom cap.
The loop is good, sterile swabs are good too. I have been using swabs recently, got a few hundred for about $5.

You're not going to collect anything from the agar, except growth to transfer from. That comes after the spores germinate.

You can flame your syringe (outside the SAB), let a drop or two fall to the SAB floor, add a drop to your swab/inoc. loop, and then start to inoculate plates. You should consider getting your agar supplies, because expensive pre-made plates add up quickly, and you're going to want at least a couple dozen to get a clean culture to grow out.
Bodhi's agar portal


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OfflineGregP
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Re: Sterilizing with Agar [Re: dankington]
    #23768595 - 10/24/16 11:08 PM (7 years, 3 months ago)

Got the prints already in Petri dishes. How long does it take plates to inoculate. Then what do I do next. That link didn't work. I have been using jars w brf and verm. Gonna get the supplies and do agar myself but I just wanted to some some before doing so. Don't have much free time on my hands for another month so I wanted to get something done then I'll learn. And learn how to do monotubs.


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Re: Sterilizing with Agar [Re: GregP]
    #23768604 - 10/24/16 11:12 PM (7 years, 3 months ago)

the link does work for me fine.
I don't know what you're talking about, so forgive me. How have you got your prints already in petri dishes?
BRF cakes? PF tek typically is just with a spore syringe. You sterilize the needle, then inoculate the jars in a SAB.

try that link again. It's good, I don't know what's up. Maybe you have new tabs blocked or something.


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OfflineGregP
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Re: Sterilizing with Agar [Re: dankington]
    #23768614 - 10/24/16 11:18 PM (7 years, 3 months ago)

I put caps over a Petri dish and spores dropped. Someone said I should clean those prints w agar?


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Invisibledankington
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Re: Sterilizing with Agar [Re: GregP]
    #23768622 - 10/24/16 11:24 PM (7 years, 3 months ago)

oh. why didn't you just make prints like normal, on foil?
Ok. Yeah. Take samples from those dishes, to your agar then. Whatever. But making prints in petri dishes is counter intuitive, and wasteful. You should really just make them on foil next time, and make agar to pour in those dishes. Instead of ordering expensive, pre-made media dishes.


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OfflineGregP
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Re: Sterilizing with Agar [Re: dankington]
    #23768633 - 10/24/16 11:28 PM (7 years, 3 months ago)

Ahhh. I see.  So swab the prints then put on the agar, then make the syringe then inoculate jars?  In a SAB?


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OfflineGregP
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Re: Sterilizing with Agar [Re: GregP]
    #23768636 - 10/24/16 11:31 PM (7 years, 3 months ago)

And how long does it take for spores to germinate. The link works now. I'll check it out tomorrow.


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Re: Sterilizing with Agar [Re: GregP]
    #23768638 - 10/24/16 11:33 PM (7 years, 3 months ago)

no. You swab to agar, and watch growth.
You sterilize a scalpel, and transfer clean looking growth to new, sterile media dishes.
Then, you take a clean wedge and either inoculate grains, or make some form of liquid inoculate or culture or whatever. Ideally, also taking a clean culture to slant for future use.


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OfflineGregP
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Re: Sterilizing with Agar [Re: dankington]
    #23768647 - 10/24/16 11:38 PM (7 years, 3 months ago)

So what size dishes should I get.


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Re: Sterilizing with Agar [Re: GregP]
    #23768656 - 10/24/16 11:43 PM (7 years, 3 months ago)

Quote:

GregP said:
So what size dishes should I get.




I use 100 mm



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Invisibledankington
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Re: Sterilizing with Agar [Re: GregP]
    #23768661 - 10/24/16 11:46 PM (7 years, 3 months ago)

get bigger ones if you can, but whatever's convenient for you.
I like 100mm dishes (actually 9~mm not exact). Bigger dishes allow you to watch growth and select clean growth. Smaller dishes don't have as much space, but they do both have advantages. Get whatever's cheapest. Get parafilm and everything too. If you have everything ready, it makes it much easier.
You can use a liquor bottle or a juice bottle to pour your media from too.


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Re: Sterilizing with Agar [Re: dankington]
    #23768677 - 10/24/16 11:54 PM (7 years, 3 months ago)

Quote:

dankington said:
get bigger ones if you can, but whatever's convenient for you.
I like 100mm dishes (actually 9~mm not exact). Bigger dishes allow you to watch growth and select clean growth. Smaller dishes don't have as much space, but they do both have advantages. Get whatever's cheapest. Get parafilm and everything too. If you have everything ready, it makes it much easier.
You can use a liquor bottle or a juice bottle to pour your media from too.





If your suggesting parafilm and the whole 9 yards you might as well get a pyrex media bottle really kicks ass


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Invisiblec10h12n2o
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Re: Sterilizing with Agar [Re: spore-ty]
    #23768723 - 10/25/16 12:33 AM (7 years, 3 months ago)

OP, you really need to read some agar basics, your questions dont even make sense, and its going to be hard for anyone to follow you since you dont know what you mean

agar is my favorite part of the hobby, i LOVE it :smile: and it is a GREAT idea to start with it from the beginning, making your own agar, pouring disposible plates, that is the short path to success. but gotta learn the basics, and read read read

its not as hard as people think, but it requires a thorough work process and good clean technique, which are skills you will pick up as you learn

best of luck!!


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OfflineGregP
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Re: Sterilizing with Agar [Re: c10h12n2o]
    #23771581 - 10/25/16 09:43 PM (7 years, 3 months ago)

Thanks all. Also, my fruiting chamber is in the closet. I always keep the door open and mist them a good deal?  Bad??  It's a big closet.



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OfflineGregP
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Re: Sterilizing with Agar [Re: GregP]
    #23771619 - 10/25/16 10:00 PM (7 years, 3 months ago)

As far as after agar, should I switch to monotubs over jars?


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Invisibler.lutece
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Re: Sterilizing with Agar [Re: GregP]
    #23771803 - 10/25/16 10:57 PM (7 years, 3 months ago)

Typically jars are used in conjunction with monotubs. Grain spawn is colonized in the jars and then spawned to CVG or manure in the monotub.


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But one does not undertake an experiment knowing one HAS failed.


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