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MagicCarpetRide89
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Registered: 12/24/11
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Agar Procedure Help!!
#16190896 - 05/06/12 03:54 PM (12 years, 8 months ago) |
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Okay, let me first start off by saying that I've been using the search bar and ALL the teks that I've been coming across have been skipping the part of pc'ing the petri dishes. And I'm not sure how I'm suppose to proceed before actually making the agar. So if anyone has a link they could send my way, I would greatly appreciate it!! (A link that has all direction from start to finish would be good, but I'll take what I can get)
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Dave_the_stoner
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I take it your using glass petri dishes, rather than presterilized disposable plastic ones? I would wrap them up in aluminum foil after lightly wetting them and sterilize them along with the agar.
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MagicCarpetRide89
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I didn't even know till just now that they sell presterilized disposable plastic ones. Had no idea. So if I had those ones, I really wouldn't need a Pressure cooker (I do plan on getting one though)?
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Doc_T
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Registered: 03/06/09
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Quote:
I'm not sure how I'm suppose to proceed before actually making the agar
is a common first step. Or were you asking a different question?
Yeah, you can buy pre-sterilized, pre-filled disposible dishes. Costs more in some ways, costs less in other ways. Lots of people use them, RR among them.
I like PF jars instead of petri dishes. Pour first, PC after. Plus, screw-on lids.
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MagicCarpetRide89
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Re: Agar Procedure Help!! [Re: Doc_T]
#16191022 - 05/06/12 04:26 PM (12 years, 8 months ago) |
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Do you have a link to a tek for that Doc_T?
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MagicCarpetRide89
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Re: Agar Procedure Help!! [Re: Doc_T]
#16191031 - 05/06/12 04:28 PM (12 years, 8 months ago) |
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Quote:
Doc_T said:
is a common first step. Or were you asking a different question?
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Doc_T
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Registered: 03/06/09
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Quote:
MagicCarpetRide89 said: Do you have a link to a tek for that Doc_T?
Search on Fast Fred's Media Cookbook for recipes. I use malt extract agar. For a liter of water, basic recipe is 30g malt, 20g agar. Use a little less agar if you're planning to start spores, it will come out softer and moister. Scale the recipe down to however dishes you want, a quarter of a batch makes about a dozen.
Add agar to cold water, stir to dissolve. Heat, stir in malt, bring to a boil, stir, turn off. It'll boil over with a quickness, don't let that happen.
That's agar solution. From there you can sterilize it and pour into sterile dishes, like RR shows in his videos. But I like to pour it into pf jars and then sterilize those and use them as dishes. 30 mins at 15 psi, then let them cool to room temp before opening. Undrilled plastic lids are my preference but grain lids can be used too.
It's hard to search for agar teks because there's several different phases. Making the dishes, pouring them, using them, the different recipes, it's a lot of info.
-------------------- You make it all possible. Doesn't it feel good?
Edited by Doc_T (05/06/12 04:41 PM)
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MagicCarpetRide89
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Re: Agar Procedure Help!! [Re: Doc_T]
#16191128 - 05/06/12 04:49 PM (12 years, 8 months ago) |
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Quote:
Doc_T said: 30 mins at 15 psi, then let them cool to room temp before opening. Undrilled plastic lids are my preference but grain lids can be used too.
Once you are ready to put your clone in, and before you open agar jar, do you put all this stuff into a glove box and do your work in there?
Quote:
Doc_T said: It's hard to search for agar teks because there's several different phases. Making the dishes, pouring them, using them, the different recipes, it's a lot of info.
I think thats way I'm having problem scaling down my search to find what I need to and read up on it.
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Doc_T
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Are you asking whether sterile work should be done in a glove box/still airbox?
Yes, is my answer to that. Unless you have a flowhood, in which case you should use a flowhood. I personally do agar work in open air on my kitchen counter. You should use a glovebox.
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MagicCarpetRide89
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Re: Agar Procedure Help!! [Re: Doc_T]
#16191228 - 05/06/12 05:16 PM (12 years, 8 months ago) |
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What kind of contam rates do you have working this way? And why not work in a glove box?
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Doc_T
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Forget I said that, use the glovebox.
Contam rates aren't really any worse than in the glove box. It's more comfortable. I do grain to grain in open air also. That's something else you should forget I said. I've got a lot of practice, I can do things smoothly and quickly enough to get away with it.
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MagicCarpetRide89
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Re: Agar Procedure Help!! [Re: Doc_T]
#16191257 - 05/06/12 05:25 PM (12 years, 8 months ago) |
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Quote:
Doc_T said: Forget I said that, use the glovebox.
Contam rates aren't really any worse than in the glove box. It's more comfortable. I do grain to grain in open air also. That's something else you should forget I said. I've got a lot of practice, I can do things smoothly and quickly enough to get away with it.
Yeah, I should probable wait tell I've got a little more experience under my belt before I venture out like that. Thanks for the help Doc_T!
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TheDeathryder
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Re: Agar Procedure Help!! [Re: Doc_T]
#16191888 - 05/06/12 07:43 PM (12 years, 8 months ago) |
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Quote:
Doc_T said: Yeah, you can buy pre-sterilized, pre-filled disposible dishes. Costs more in some ways, costs less in other ways. Lots of people use them, RR among them.
I like PF jars instead of petri dishes. Pour first, PC after. Plus, screw-on lids.
I've been trying to find rhese plastic pre filled agar dishes since you mentioned them, and have had no luck. Does anyone know where to find them?
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Doc_T
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-------------------- You make it all possible. Doesn't it feel good?
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MagicCarpetRide89
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Re: Agar Procedure Help!! [Re: Doc_T]
#16191915 - 05/06/12 07:48 PM (12 years, 8 months ago) |
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Quote:
Doc_T said: Try this link.
Thanks Doc_T (even though this wasn't really for me). btw Doc_T what is it that girl is staring at??
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TheDeathryder
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Re: Agar Procedure Help!! [Re: Doc_T]
#16191945 - 05/06/12 07:54 PM (12 years, 8 months ago) |
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Quote:
Doc_T said: Try this link.
I googled is already before posting, got results for powdered agar and petri dishes of sheeps blood agar whatever the hell that is.
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Doc_T
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Registered: 03/06/09
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Second link on that page gets me bulk lots of blank dishes. I'll leave the rest as an exercise for the student. If you can't find and buy disposable petri dishes you aren't ready to have them yet and Google knows it.
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MagicCarpetRide89
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Re: Agar Procedure Help!! [Re: Doc_T]
#16191995 - 05/06/12 08:06 PM (12 years, 8 months ago) |
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Damn....I've been sooo lazy today. I should of googled that. And there's really no excuse for me not being able to do that
I've only been sitting at the computer ALL damn day. You've been so helpful too Doc_T And how does one accumulate all those post in three years?? 35 thousand some post is a lot sir.
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spornication
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you can reuse your plastic petri dishes with a sanitary dillution of bleach should never buy pre poured dishes your just asking for contams but they are cheap i get 20 for 3.00us on ebay there is a bid on 100 right now, get on it 3 days left ive got enough i think the bid is at 12.00 at this moment
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Edited by spornication (05/06/12 11:12 PM)
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IGetTight
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Doc you've been changin that boxxy pic a lot lately. I never really realized how hot she was. On a side note I've got everything i need for agar work and still have not created an isolate.... slackin hard!
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joyless
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Quote:
spornication said: you can reuse your plastic petri dishes with a sanitary dillution of bleach should never buy pre poured dishes your just asking for contams
thats BS. where i buy my pre poured plates, they come vaacum sealed with lab quality check and bacterial count.
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MagicCarpetRide89
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Re: Agar Procedure Help!! [Re: Doc_T]
#16200882 - 05/08/12 05:49 PM (12 years, 8 months ago) |
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Should I clone from my forth flush? Or would that not work so good?
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Edited by MagicCarpetRide89 (05/08/12 07:23 PM)
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MagicCarpetRide89
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....bump.....
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bishlap
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it should be fine but I would isolate from it asap then inoc a few more plates and get your isolate if you didn't already.
time I assume would be your enemy because of senescence, were they from g2g?
-------------------- "If you're not worried that you took way
to much, you didn't take enough" -
Terrence McKenna
There is no soul, only the ego dies.
The body was never yours.
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MagicCarpetRide89
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Re: Agar Procedure Help!! [Re: bishlap]
#16206202 - 05/09/12 06:21 PM (12 years, 8 months ago) |
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No, MS. But the first 3 flushes were real nice. I wish I cloned from one of those first three. You think even if I clone from this last flush, I will still be isolating a good clone?
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bishlap
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the sample will likely have many "strains" in it. I'm assuming you want to save your isolates for later use in slants so if that's the case you'll want to try to get your isolates taken as soon as you see sectoring to multiple other dishes from each healthy sector
grow those till/if they sector and isolate again if needed, place samples in slants, grow out each isolate and decide what one you like best then use that slant to colonize more agar > grain > g2g > whatever floats your boat
remember to label everything, or life will brcome difficult for you later.
the only issue I can think of from cloning soo late is senescence weaking your culture but you should be fine if your diligent.
-------------------- "If you're not worried that you took way
to much, you didn't take enough" -
Terrence McKenna
There is no soul, only the ego dies.
The body was never yours.
Edited by bishlap (05/09/12 07:05 PM)
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