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BlueIndian
Maestro



Registered: 01/17/10
Posts: 858
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Newbie agar questions
#17840812 - 02/21/13 11:16 AM (11 years, 2 months ago) |
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I'd like to start working with agar but not sure I can get it right using a glovebox? I don't have funds to build a flowhood right now. If I obtained pre-made agar plates is it ok to work with agar in a glovebox or is there too much potential for failure? And how long can agar plates be stored in the fridge? Is there a difference in shelf life of just agar or agar that has a culture on it? I ask because if I bought say 20 plates and just used 5-6 to begin with it may be a couple of months before I would use the others. I wouldn't dare try to pour agar in the open where I am and it seems way too awkward to try to pour in a box. I'd rather just pay a few bucks each and be done with it.
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entactogenics
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Re: Newbie agar questions [Re: BlueIndian]
#17840850 - 02/21/13 11:24 AM (11 years, 2 months ago) |
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ive just started as well, so im not the definitive thats for sure. i ordered ten plates from outgrow and used them over a period of 2 months, storing them at room temp. they seemed to keep fine, and i only used a still air box, and im sure my sterility isnt great. the last plate is colonizing grain now and looking awesome. ten for ten, not bad for a newb.hopefully this inspires you to take the leap.
thhis isnt really adv myco for future reference. mush cult would make more sense
Edited by entactogenics (02/21/13 11:26 AM)
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PussyFart
Retired Cultivation Extrodinaire



Registered: 04/08/12
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Re: Newbie agar questions [Re: BlueIndian]
#17840864 - 02/21/13 11:28 AM (11 years, 2 months ago) |
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A still air box is more practical for this hobby than a glove box, the only difference in the 2 being the attached gloves.
I do all of my agar work in a SAB, it is the next best thing to a flow hood, and there is nothing in this hobby that cannot be done in one.
Agar plates, in the fridge, a couple months maybe, but master slants in the fridge, years.
I store myt unused dishes at room temp for months, and then in the fridge when I want it to stall for a bit with a culture.
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THIS HOBBY IS NOT FOR THE IMPATIENT! PLEASE BE PATIENT, DON'T BE A PATIENT! A Tale of 10 Isolates, GT Cluster Clone Monotubs, RR's Let's Grow Mushrooms DVD, SGFC(Shotgun Fruiting Chamber), Monotub Tek, Damion5050's Coir Tek, TL's Tek List, Frank's Tek List, EvilMushroom666's Pasteurization Tek, How It Should & Shouldn't Look - NEW CULTIVATORS GUIDE *** *** AFGHAN KUSH GROW LOG *** ***
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BlueIndian
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Registered: 01/17/10
Posts: 858
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Re: Newbie agar questions [Re: PussyFart]
#17842148 - 02/21/13 02:51 PM (11 years, 2 months ago) |
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Awesome. Thanks for the replys! I do have the ability to remove my gloves and use it as a SAB so that might make it easier. Probably be able to work faster without being hampered by the gloves. I'm just looking to be able to do some isolating and working with cloning for now. If I can have success this way then I can try to make slants for longer storage. Multi spores just aren't the way to go so this is what I was hoping for. Thanks guys!
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thiotimoline
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Registered: 12/01/12
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Re: Newbie agar questions [Re: BlueIndian]
#17844843 - 02/21/13 10:32 PM (11 years, 2 months ago) |
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Pouring agar in a still air box is not too hard. Just make sure the box is tall enough.
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HIGHIMTHENEWGUY


Registered: 05/15/08
Posts: 162
Loc: Wonderland
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Re: Newbie agar questions [Re: BlueIndian]
#17845142 - 02/21/13 11:55 PM (11 years, 2 months ago) |
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I completed my Flowhood over summer. Prior to that, I did everything in a SAB. Like others said just make sure it is tall enough. Most people are usually pouring out of empty liquor/beer bottles. I will say it is definitely way more comfy outside of one and using a flowhood, but assuming your sterile technique is proper i see no reason why you can't achieve a minimum of 80% success rate for clean plates. The beauty of agar: If your plate does develop a contaminant, you get to transfer to a new plate. Once you start using agar you may never go back to doing LC's or MS.
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Terry M
Stranger in a Strange Land



Registered: 06/18/10
Posts: 1,502
Loc: Rhode Island
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Around here, we use the terms "glove box" and "still air box" interchangeably. But what we nearly always mean is "still air box," with no attached gloves. The latter is too clumsy for mycology.
-------------------- Liberté, égalité, humidité.
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BlueIndian
Maestro



Registered: 01/17/10
Posts: 858
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Re: Newbie agar questions [Re: Terry M]
#17846670 - 02/22/13 10:59 AM (11 years, 2 months ago) |
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I'm going to give it a shot. I salvaged a clear acrylic reptile habitat box from the side of the road and cut out arm holes. (yeah I cleaned it out very well). Nice thing about it is it's easy to see through and has good height to it. Hopefully I can find some agar at one of the local grocery stores...if not we have a few asian specialty stores. I like the potato/agar/honey recipe. It does state brewers yeast is optional. How much difference will omitting the yeast cause? There is a brew supply place within 4 minute drive from me.
Also looking for some petri dishes. One place has 60mm x 15mm (20 pack) for $10 shipped or they have 100mm x 15mm (20 pack) for $14 shipped. Which size would be the better option? Again thanks for everyones help! I'm so ready to bust away from the multi spore route.
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PussyFart
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Registered: 04/08/12
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Re: Newbie agar questions [Re: BlueIndian]
#17846761 - 02/22/13 11:17 AM (11 years, 2 months ago) |
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Quote:
BlueIndian said: Also looking for some petri dishes. One place has 60mm x 15mm (20 pack) for $10 shipped or they have 100mm x 15mm (20 pack) for $14 shipped. Which size would be the better option? Again thanks for everyones help! I'm so ready to bust away from the multi spore route.
A case of 500 for about $70 seems like the deal.
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THIS HOBBY IS NOT FOR THE IMPATIENT! PLEASE BE PATIENT, DON'T BE A PATIENT! A Tale of 10 Isolates, GT Cluster Clone Monotubs, RR's Let's Grow Mushrooms DVD, SGFC(Shotgun Fruiting Chamber), Monotub Tek, Damion5050's Coir Tek, TL's Tek List, Frank's Tek List, EvilMushroom666's Pasteurization Tek, How It Should & Shouldn't Look - NEW CULTIVATORS GUIDE *** *** AFGHAN KUSH GROW LOG *** ***
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BlueIndian
Maestro



Registered: 01/17/10
Posts: 858
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Re: Newbie agar questions [Re: PussyFart]
#17846865 - 02/22/13 11:37 AM (11 years, 2 months ago) |
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500 is a little overkill for me I think. Hoping success rate will be higher than that....don't see going through 20 in a year really. Also I read to use empty liquor bottle or similar to mix and PC. Wouldn't a clean quart jar with foil on top work just as good or would it be messy trying to pour from it? If it comes down to it I guess I can finish off a bottle in the cabinet...if I REALLY have too...
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PussyFart
Retired Cultivation Extrodinaire



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Posts: 22,502
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Re: Newbie agar questions [Re: BlueIndian]
#17846932 - 02/22/13 11:53 AM (11 years, 2 months ago) |
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A bottle with a long neck is key, the long neck makes it easier to pour than a jar, and the small opening allows for less contams to wander in.
Success rate has little to do with it, If you have one culture and make a few transfers from one dish, that is about 5 dishes, but then take transfers from them and your 20 are gone...so they do kinda go fast, but I guess it all depends on what your plans are.
If you plan on isolating and ccloning strains, I suggest you stock up on them, rather than buy them by the sleeve and make someone else rich.
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THIS HOBBY IS NOT FOR THE IMPATIENT! PLEASE BE PATIENT, DON'T BE A PATIENT! A Tale of 10 Isolates, GT Cluster Clone Monotubs, RR's Let's Grow Mushrooms DVD, SGFC(Shotgun Fruiting Chamber), Monotub Tek, Damion5050's Coir Tek, TL's Tek List, Frank's Tek List, EvilMushroom666's Pasteurization Tek, How It Should & Shouldn't Look - NEW CULTIVATORS GUIDE *** *** AFGHAN KUSH GROW LOG *** ***
Edited by PussyFart (02/22/13 11:53 AM)
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HIGHIMTHENEWGUY


Registered: 05/15/08
Posts: 162
Loc: Wonderland
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Re: Newbie agar questions [Re: BlueIndian]
#17849913 - 02/22/13 09:55 PM (11 years, 2 months ago) |
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At one point I thought the same thing as you. Once you start getting into agar work though you will realize how fast you go through plates. I am working on only two different isolations right now and am going through alot of plates. I would just buy a case of 500 or at minimum half that. If your only out to make one isolate you could do it within a single sleeve, but thats not leaving you a whole lotta wiggle room in the chance of error or if you aren't familiar with the process. It really all depends about how deep your into this hobby I guess. Over time for me it has grown into a passion, a labor of love, and something I find myself obsessing over in my spare time. I think alot of members here have mutual feelings towards this hobby
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BlueIndian
Maestro



Registered: 01/17/10
Posts: 858
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Well that was an interesting adventure. Went to the so called 'super grocery store' and no luck. So I went to an oriental import place that has karate gear, throwing stars, rice cookers...all that crap. Walked in and heads turned like I was from another planet. "Mr. Li" says yes the have agar...go to the back of the store and go down stairs. I'd been in there once before and didn't even know they had a downstairs. So...walk down there and there are aisles of every damn noodle you could want. Cans and boxes of stuff I had no idea what it was. Freezers full of god knows what...fermented duck eggs etc. Bags and bags of dried oyster mushrooms. I hunted all over and had to ask several Koreans in there if they knew where the agar was. No one knew. Finally Mr. Li comes down and shows me...
So what they had were the long bars and some were green or red. I thought that would look cool but looked and those had x amount of sugar in them and I didn't know if that was good or bad. Didn't have any clear so a yellowish one was the only one I saw with just agar. Got 2 of those...see if I can get a pic here.
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BlueIndian
Maestro



Registered: 01/17/10
Posts: 858
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Re: Newbie agar questions [Re: BlueIndian]
#17852866 - 02/23/13 02:40 PM (11 years, 2 months ago) |
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So before I found those pieces I had a box in my hand of flakes. I went ahead and bought it too but the first ingredient listed is sugar and then agar. Is this usable? The recipe I was looking at is potato water/agar/honey. If I used these flakes would that eliminate the need for honey? I just got what I got and got out of there. I'm ready to do this...just have to get the petris now.
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Gewehr
The Gun



Registered: 05/18/12 
Posts: 12
Loc: Pacific Northwest
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Re: Newbie agar questions [Re: BlueIndian]
#17858317 - 02/24/13 03:08 PM (11 years, 2 months ago) |
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It looks like what you have picked up is food grade "agar" with sugar in it. Those products are more like gelatin. Biological grade agar generally comes as a powder and you add water to it. Even better, you can order potato dextrose agar online from any mycology supply store and skip a lot of steps as it already has everything in it that you need. Also, pouring plates in a SAB is probably not a bad idea, but not really necessary. I pour plates for my job at a university biology department almost everyday and we always pour them in open air. You just need to make sure your pouring technique is good and quick, and that you are in a room with little air movement.
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Jeff
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Re: Newbie agar questions [Re: Gewehr]
#17858419 - 02/24/13 03:32 PM (11 years, 2 months ago) |
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There are some "food grade" agars that don't have additives. It is hard to tell sugar content from an ingredient list on required labeling. I would not even know where to begin with mix %. Some of the vendors listed here have small quantities that you can pick up at a reasonable price.
-------------------- Myco-tek
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Terry M
Stranger in a Strange Land



Registered: 06/18/10
Posts: 1,502
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Re: Newbie agar questions [Re: Gewehr]
#17858426 - 02/24/13 03:34 PM (11 years, 2 months ago) |
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Quote:
Gewehr said: Also, pouring plates in a SAB is probably not a bad idea, but not really necessary. I pour plates for my job at a university biology department almost everyday and we always pour them in open air. You just need to make sure your pouring technique is good and quick, and that you are in a room with little air movement.
??? How long are these plates good for? How long do your plates last before contamination starts growing? For mycology, it should be "never." My plates, properly poured under sterile conditions, simply do not contaminate unless I make a slip-up pouring. They can go months at room temperature, until the agar gets dry and starts shriveling up.
-------------------- Liberté, égalité, humidité.
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Gewehr
The Gun



Registered: 05/18/12 
Posts: 12
Loc: Pacific Northwest
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Re: Newbie agar questions [Re: Terry M]
#17858605 - 02/24/13 04:18 PM (11 years, 2 months ago) |
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They don't contam.. Otherwise why would we pour them like that? I'm saying its generally not the air that is the problem it pouring methods. e.g. condensation wicking in contaminants or leaving an agar bridge on the edge of the plate. At most we may get 1 or 2 plates that contam, if any. But we are pouring hundreds of plates a day. If you are good and quick, you are introducing your sterile plate to open air for a very short amount of time and the only thing that is ever above it should be its lid and the sterile tip of your pour bottle.
Subsequent transfers and other procedures should be done in a sab or in front of a LFH because you are no longer dealing with things that are perfectly sterile. i.e. the outside of the petri dish has been sitting exposed to air instead of resting inside a sterile sleeve. You are also now moving objects over the open plate much more which is the largest source for contams.
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BlueIndian
Maestro



Registered: 01/17/10
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Re: Newbie agar questions [Re: Gewehr]
#17858726 - 02/24/13 04:44 PM (11 years, 2 months ago) |
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Ok well i did a search for teks and came up with the grocery store agar tek. The box of powder has sugar listed as first ingredient so maybe it isn't what I want. The yellow bar things are labeled Agar-Agar (gulaman). Product of the Philippines. Ingredients listed are seaweed, FD&C yellow #5. Plans were to mix this with potato water and honey and maybe some brewers malt. Is this not going to work? If not it's $6 lost but it's the time and effort later that I'd rather not expend energy on if it won't work. Just tried to go the cheap route and most times that costs you more in the long run...
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RogerRabbit
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Re: Newbie agar questions [Re: Gewehr]
#17860218 - 02/24/13 08:51 PM (11 years, 2 months ago) |
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Quote:
Gewehr said: They don't contam.. Otherwise why would we pour them like that? I'm saying its generally not the air that is the problem it pouring methods. e.g. condensation wicking in contaminants or leaving an agar bridge on the edge of the plate. At most we may get 1 or 2 plates that contam, if any. But we are pouring hundreds of plates a day. If you are good and quick, you are introducing your sterile plate to open air for a very short amount of time and the only thing that is ever above it should be its lid and the sterile tip of your pour bottle.
Subsequent transfers and other procedures should be done in a sab or in front of a LFH because you are no longer dealing with things that are perfectly sterile. i.e. the outside of the petri dish has been sitting exposed to air instead of resting inside a sterile sleeve. You are also now moving objects over the open plate much more which is the largest source for contams.
Medical testing labs get away with this sort of sloppy technique because they're usually growing out bacteria cultures and then disposing of them a few days thereafter. They also use nutrient agar, not MEA so it doesn't grow molds as well. If you poured MEA or similar in open air it would be green in a few days. 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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