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Josborne
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Why agar transfers getting weaker? 1
#27308433 - 05/14/21 01:46 PM (3 years, 8 months ago) |
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I've been doing multiple transfers from the same culture and the last batch all came out looking like the bottom out (good growth in some places, weak looking growth in others) even though the master plate looked like the top one (uniform dense growth).
Many of the plates in the new batch look even worse than the bottom one pictured with all/most of the growth is very weak/whispy looking
Is this something to be worried about?
Edited by Josborne (05/14/21 01:47 PM)
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seldom seen
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Re: Why agar transfers getting weaker? [Re: Josborne]
#27308442 - 05/14/21 01:51 PM (3 years, 8 months ago) |
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How many transfers? Maybe switch up the media?
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Josborne
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Re: Why agar transfers getting weaker? [Re: seldom seen]
#27308490 - 05/14/21 02:20 PM (3 years, 8 months ago) |
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Quote:
seldom seen said: How many transfers? Maybe switch up the media?
This is maybe the fifth "batch?" What I've been doing is I transferred from the best looking master plate into 10-15 or so receiving plates. Then the same for the best looking plate of the second batch. And so on. So effectively I've done 70 or so plates from one original master plate/culture.
Do you mean changing agar to water ratios or using something besides agar? What do you use?
Edited by Josborne (05/14/21 02:22 PM)
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Josborne
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Re: Why agar transfers getting weaker? [Re: Josborne]
#27308500 - 05/14/21 02:27 PM (3 years, 8 months ago) |
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Also at first I thought this might be senescence, but after a little reading it seems like the consensus on this forum is that senescence takes a long long time to happen and is not understood very well anyway.
Is that green light to transfer, clone, g2g, fruit, etc as much as I want? Or will I eventually have to go back to spores? (speaking within a year timeframe)
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sandman420
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Re: Why agar transfers getting weaker? [Re: Josborne] 1
#27308517 - 05/14/21 02:37 PM (3 years, 8 months ago) |
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How big of a batch of agar do you make at one time? (ml)
A possible explanation could be that less genetics per transfer equals less dense growth.
Or maybe you messed up your agar recipe last round and that made a lot of weak plates possibly?
I mean both of those plates look just fine to me. But you said you had a whole batch of whispy ones so...
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smalltalk_canceled
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Re: Why agar transfers getting weaker? [Re: sandman420]
#27308536 - 05/14/21 02:45 PM (3 years, 8 months ago) |
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Why is the bottom plate not good? Weak growth what
-------------------- Willpower is the one true virtue
  
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Josborne
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Re: Why agar transfers getting weaker? [Re: sandman420]
#27308540 - 05/14/21 02:46 PM (3 years, 8 months ago) |
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Quote:
sandman420 said: How big of a batch of agar do you make at one time? (ml)
A possible explanation could be that less genetics per transfer equals less dense growth.
Or maybe you messed up your agar recipe last round and that made a lot of weak plates possibly?
I mean both of those plates look just fine to me. But you said you had a whole batch of whispy ones so...
I usually do 250ml. (250ml water/12g malt extract agar)
What kind of mess up would contribute to weaker plates? Is extra agar powder worse than less?
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Josborne
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Quote:
smalltalk_canceled said: Why is the bottom plate not good? Weak growth what
It's not as uniform/dense looking as my previous plates. The 9-12 o clock area looks a lot weaker than the rest of the plate. But I might be totally over thinking it haha. I was just surprised at the difference because my plates thus far have been pretty consistent.
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sandman420
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Re: Why agar transfers getting weaker? [Re: Josborne]
#27308556 - 05/14/21 02:58 PM (3 years, 8 months ago) |
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When you do an extra small batch of agar that means that any mistakes or errors in the recipe are multiplied.
250 agar recipe: 5g agar 5g malt. 6 grams is 20% overshot. 7 grams is 40% overshot. 3 grams is 40% undershot. You get what I'm saying? How accurate is your scale even? I really recommend to make bigger batches, at least 500ml.
Edited by sandman420 (05/14/21 02:59 PM)
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Josborne
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Re: Why agar transfers getting weaker? [Re: sandman420]
#27308570 - 05/14/21 03:11 PM (3 years, 8 months ago) |
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Quote:
sandman420 said: When you do an extra small batch of agar that means that any mistakes or errors in the recipe are multiplied.
250 agar recipe: 5g agar 5g malt. 6 grams is 20% overshot. 7 grams is 40% overshot. 3 grams is 40% undershot. You get what I'm saying? How accurate is your scale even? I really recommend to make bigger batches, at least 500ml.
Oh wow. I never even thought of that. Thanks for pointing that out! 
And it's a pretty cheap scale so I'm assuming not crazy accurate. I will take your advice.
I originally was doing 500ML but found I always had a lot of extra left over unless I did at least 30 plates at one time—which it sounds like I might have to start doing!
I just hate wrapping plates. If I don't transfer during that session then it means I'll have to parafilm twice (once after solidified, then again next time I open it for a transfer). However I saw that some people just wrap the petri dish bag over a stack of plates until they decide to use them so I might do that...
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sandman420
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Re: Why agar transfers getting weaker? [Re: Josborne]
#27308589 - 05/14/21 03:24 PM (3 years, 7 months ago) |
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I put them in stacks in large 2 gallon ziplocs straight in the hood, but I don't have sleeves because glass petris.
When I used sleeves I would take a few out so I could tuck the bottom all snug because I'm a pervert. Put those in another ziplock or suomething.
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Shakedown Street
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Re: Why agar transfers getting weaker? [Re: sandman420]
#27309081 - 05/14/21 09:40 PM (3 years, 7 months ago) |
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During winter past, I got good at making agar. So I decided to have a speed race. I made an inventory of all the possible spore prints, petri dishes of agar, and jars of LC I had that might possibly grow mycelium. Then I made that number of agar petri dishes and inoculated each and every one, marking each petri dish carefully to denote its source.
My only goal was to determine which agar made it to the edge of the petri dish first, #1,and which came in second place, #2.
Thereafter, I only transferred winners #1 and #2 to grain. My reasoning was that I only wanted the mycelium that most quickly colonized agar because that mycelium would also most likely colonize grain the quickest, and avoid contamination.
My speed race was just a farmer's version of Darwin: save the best, trash the rest.
Remember all life depends on water, and microbes need it too. The faster the mycelium sucks it all up, the less likely some stray spore or bacterium can grow. That was my reasoning.
Another positive thing that came from this speed race was that I easily trashed a whole shitload of losers.
After doing this I learned G2G transfers, a technique that allows you to turn one fully-colonized grain jar into ten fully colonized grain jars in 10 to 14 days. Thanks to PhillyGoldenTeacher.
Because of this, I have many, many fully colonized grain jars. Only one or two came out duds--that is, only a single clump mycelium growing in them. Straight to the compost heap they went.
I recall your darkness when it crackled like a thundercloud.
Edited by Shakedown Street (05/14/21 09:47 PM)
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