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jordanhess0319
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Registered: 03/20/16
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Last seen: 4 years, 10 months
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agar
#23395361 - 06/29/16 08:00 PM (7 years, 6 months ago) |
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Anyone ever use this premixed agar ? how were the results, and how did you mix it im not having much luck with it

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weetsie
unlicensed tub surgeon



Registered: 05/08/11
Posts: 572
Loc: United Kingdom
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In what way are you not having much luck?
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cronicr



Registered: 08/07/11
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works great, make sure to start with cold water or it clumps.
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jordanhess0319
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i hope all of this makes sense
ive tried taking a few clones on the agar and they either dont take or the growth is really slow, and i tested some liquid culture and most of them never grew the few that did tho, the growth was really whispy and seemed super slow.
on the container the premix came in it says to mix the whole 100 grams with 1800 ml water, but i wanted to make a smaller batch so i mixed it 4 grams to 100 ml water then heated the water up to a boil on the stove like it says on the container and then put it in my pouring bottle and pressure cooked for 15 min. at 15 psi.
then i read that agar is just like liquid culture in the fact that it is better to have less nutrients rather than to many. so i made a batch that was 3.5 grams to 100 ml water and did everything else the same way. had pretty much the same results with this mixture.
i hope the pics are good enough to get a general idea of what im workin with
the dish on top is one of the liquid cultures i tested and i thought it looked good so i made a few transfers from the dish, and hopefully from the dates you can tell whats goin on #1

the top one here are transfers from a clone and the bottom one is a transfer from one of the top dishes - seems really really slow to me #2

top dish here is a transfer from a clone thats like a month old and the bottom one is a 8 day old transfer from a dish i tested a king oyster liquid culture on and there is no growth #3

here is the first transfer and second transfer from a king oyster dish i tested a liquid culture on the top dish here looks more like pictures i see, but still seems like slow growth #4

maybe my powder to water ratio is wrong, or not enough nutrients, to many nutrients bad procedure, maybe its all normal i dont really know im new with agar
Edited by jordanhess0319 (06/29/16 10:56 PM)
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weetsie
unlicensed tub surgeon



Registered: 05/08/11
Posts: 572
Loc: United Kingdom
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Something isnt right, oysters take about 6-9 days to cover a 90mm plate using MEA.
If you're confident your cultures are good I'd suggest trying some new agar, buy an oz for a couple of $ and you can rule out your premixed MEA as the problem if you don't get better results.
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jordanhess0319
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Registered: 03/20/16
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Last seen: 4 years, 10 months
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im not confident in my cultures yet but i thought they looked good as in no contaminations, other people have said they have used it with good results idk what im doin wrong. maybe ill do like you suggested and just try another kind. thanks for your input.
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jordanhess0319
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Quote:
cronicr said: works great, make sure to start with cold water or it clumps.
how much powder and water did you use in the mixture ? maybe im just mixing it wrong
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Perception7
Psilocin Technician



Registered: 01/10/16
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Edited by Perception7 (06/30/16 03:52 PM)
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jordanhess0319
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Last seen: 4 years, 10 months
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on the container it says to mix the whole container (100 grams) with 2500 mls water so i just scaled it down so i could do a 100 ml batch so i put in 4 grams of the powder. and thats the agar, potato flakes and everything mixed together already.
this is what munchauzen said he uses and i assume it works for him,
For 8-10 Pasty Plates You Will Need: - 1 gram honey or raw sugar - 2 grams agar - 3 grams potato flakes - 1/2 cup water - optional: food coloring
1/2 cup water measures out to roughly 125 ml so everything added together in his recipe weighs 6 grams and thats allot more than what ive been mixing in mine so maybe if i added 2 - 3 grams of potato flakes along with the four grams of premix it would work.
maybe there isnt enough nutrients in this brand of premix is what im thinking
does this make sense ?
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mupetmower
Mower of Muppets



Registered: 03/29/16
Posts: 3,036
Loc: Here and There
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i actually use less potato flakes than agar, and mine comes out alright.
i use the recipe in Pasty's Pasty Plate tek.
-------------------- -The wise man never stops seeking knowledge.
-I wanna feel the change consume me, feel the outside turning in. I wanna feel the metamorphosis and cleansing I've endured within.
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SpitballJedi
Ancient Astronaut



Registered: 10/13/12
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Loc: Nibiru
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Quote:
jordanhess0319 said: Anyone ever use this premixed agar ? how were the results, and how did you mix it im not having much luck with it


Your agar looks a lot lighter and clearer than mine does, but I don't use that pre-made MAE.
Says mix 100g MAE with 1.8L water.
1.8L=1800mL
Since you used 100mL water, you divide 100g by 18.
if you are only using 100L water, then you use 5.55g of the MAE. You shouldn't have to mix anything else with it.
Since you only used 4g, that might not be rich or thick enough. Since I've never used it, I can't say for sure, but according to the directions, you didn't use enough.
Math.
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jordanhess0319
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for the mea premix i mixed 6 grams with 108 ml water and then for the pda premix i used 4 grams with 100 ml water i think my math was good i just screwed up how i typed it in the post - i do suck at math tho
i just made a batch of pda, this time i used 4 grams of premix and 3 grams potato flakes with 100 ml water and just poured 5 plates
next im going to make a batch of mea and use 7 grams of premix with 100 ml water and pour 5 plates
maybe i just mixed both kinds a little to weak, i don't know.... i dont have the money to buy different agar right now so im just going to experiment with different ratios i think and see if anything happens.....im so frustrated i just wasted 8 half gallon jars and 10 quart jars of rye due to bacteria in a liquid culture, so i figured id learn agar so i can guarantee a clean culture and now i cant get the fricken agar to work right
Edited by jordanhess0319 (07/01/16 11:50 AM)
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mupetmower
Mower of Muppets



Registered: 03/29/16
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even with those ratios, it still seems a little strange to get almost 0 growth. i mean, i have made my PDA with lots of different ratios, trying to get a feel for how much makes it thick and how much i need for softer, etc. off all the times, i have only 1 times not gotten ANY growth, and that was with an AA+ syringe from a vendor that was giving me trouble everywhere.
If it gels up like agar should, then it seems like it should grow, even if maybe a little slow or something. you know? idk im not an expert, so idk. and i have only ever made PDA, with powdered agar, potato flakes, water, and drop of karo. never made any premix stuff. just seems odd. maybe it was the tissue sample you took, or something.. where the fruits like super mature or anything? and as for the LC, did it grow any contams or anything?
EDIT - sorry, just went back and read one of your earlier posts again, saying you did get growth, just some slow and wispy.
hmm.. still just seems odd to have that much trouble with it, with just having your ratio a little off. are you certain it couldnt be the tissue sample or the LC or something?
EDIT2 - also, that LC plate you took a transfer from, didnt seem to have a lot of strong growth on it, when you transferred. as for the clone transfers, i noticed that when i took some transfers from a more mature clone plate, it had really wispy growth that grew kinda slow, kinda like youre talking about. so maybe it is more the cultures and stuff you are using rather than the agar ratio. im not 100% sure, just making an observation.
hope you get it worked out.
-------------------- -The wise man never stops seeking knowledge.
-I wanna feel the change consume me, feel the outside turning in. I wanna feel the metamorphosis and cleansing I've endured within.
Edited by mupetmower (07/01/16 10:57 AM)
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jordanhess0319
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Registered: 03/20/16
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yea i hope i get it worked out to, im sure i will it just takes some learning and experience it is frustrating sometimes tho......not gonna give up tho bc i love this hobby
thanks everybody for the input
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jordanhess0319
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Registered: 03/20/16
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Last seen: 4 years, 10 months
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  got some good growth from that last batch i made i added potato flakes to the PDA premix and i just added 2 grams more of the powder to the MEA premix per 100 ml water
i think the problem was the cultures tho bc i did a few transfers on these new plates and got good growth and i also did some transfers to some plates i poured a couple weeks ago with the first ratio i tried and got good growth....is that what people mean when they say "do a few transfers to clean up the culture"
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weetsie
unlicensed tub surgeon



Registered: 05/08/11
Posts: 572
Loc: United Kingdom
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Quote:
jordanhess0319 said:
  got some good growth from that last batch i made i added potato flakes to the PDA premix and i just added 2 grams more of the powder to the MEA premix per 100 ml water
i think the problem was the cultures tho bc i did a few transfers on these new plates and got good growth and i also did some transfers to some plates i poured a couple weeks ago with the first ratio i tried and got good growth....is that what people mean when they say "do a few transfers to clean up the culture"
You're probably right about your cultures being dirty.
From what I can see the plates look good, if they were mine the next transfer would be to grain
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