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Bertoxulous
Stranger

Registered: 04/26/16
Posts: 23
Last seen: 8 years, 8 months
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1st ever Agar Inoculation, Squirt or Swab?
#23194573 - 05/06/16 05:57 AM (8 years, 8 months ago) |
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I'm doing my first ever agar inoculation today and I was curious what the best method would be for transferring from a syringe.
I've heard sterilized swabs mentioned before, and I do have some that I made. However, I see this method as an added contam factor.
I'm sure there are benefits to squirting some spore solution on a swab and then wiping onto the plate. Such as, no water sitting on the plate after the inoculation, control over the inoculation points on the plate.
I do not plan on doing any isolation. My goal is to simply ensure that I have a clean specimen to work with, and then transfer from agar to grain.
So the question is, should I squirt or swab?
Also, some explanation behind the steps taken in either method to ensure sterile technique would be appreciated, I know the basics, but I'm still brand new to agar.
If it's recommended that I squirt the solution, should I hold the agar on an angle, letting the solution drop down it, and then perhaps poor off the excess water?
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Supalemonhaze
Spore syringe hater.



Registered: 10/02/15
Posts: 6,725
Loc: 12" down Europe's butthole
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Re: 1st ever Agar Inoculation, Squirt or Swab? [Re: Bertoxulous]
#23194588 - 05/06/16 06:10 AM (8 years, 8 months ago) |
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Anywhere from 1-3 drops is good enough and there won't be much water running around the plate after a few days due to evaporation so you don't have to drain it. You would probably dump most of the spores with it anyway.
I did try swabbing once with spore solutions but the vial I was using was not viable so I got no germination. Spore solutions can be a bit of a headache to germinate on agar, I have a pan spore solution which is being stubborn.
If you ever have problems with syringes on agar, you should try it with grains or even better, an LC. Then you can fish out the grains/mycelium that germinates and put that on agar. I inoculated 3 lcs yesterday with pans since I haven't seen any results on agar so far.
I use swabs on PE only, those do not like to drop spores so swabbing is one of the only ways to store spores.
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Mad Season
hookers and blackjack



Registered: 09/16/12
Posts: 12,666
Loc: Canada
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Re: 1st ever Agar Inoculation, Squirt or Swab? [Re: Supalemonhaze]
#23194601 - 05/06/16 06:18 AM (8 years, 8 months ago) |
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ya know making a softer more nutritious agar would accomplish what LC and spawn does.
There's also forced hydration, hot pour, and charcoal agar to help with germination.
Agar is the king for germination. The only unviable culture I couldn't get to germinate on agar, also wouldn't germinate in spawn.
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Supalemonhaze
Spore syringe hater.



Registered: 10/02/15
Posts: 6,725
Loc: 12" down Europe's butthole
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Re: 1st ever Agar Inoculation, Squirt or Swab? [Re: Mad Season]
#23194617 - 05/06/16 06:35 AM (8 years, 8 months ago) |
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My Galindoi vial was like that, made a bunch of plates and warm poured them all after a couple weeks. After I gave up on it I used it all on LCs and grains. None got germination.
I have used this pan vial on soft and normal agar so far excluding the LCS I inoculated yesterday. I skipped the warm pour for this vial because I find it annoying. An LC is fast and easy and will give you results right away, unlike testing agar methods. IMO if spores won't germinate in an LC, they won't do on anything else. I even prefer it more than spores to grain.
One question though, isn't forced hydration useless with spore solutions? I thought that only applied for old prints.
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Mad Season
hookers and blackjack



Registered: 09/16/12
Posts: 12,666
Loc: Canada
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Re: 1st ever Agar Inoculation, Squirt or Swab? [Re: Supalemonhaze]
#23194637 - 05/06/16 06:49 AM (8 years, 8 months ago) |
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I'd really recommend trying activated charcoal agar. Spore syringe vendors put a bit of charcoal into the syringes to help germinate spores too. Lots of the time that's what the black specks are in the solution lol
Forced hydration works for old spores in a syringe too. I got a couple PE syringes I couldn't get to germinate until I did the forced hydration tek
I also heard snake venom can help too, but who has that right? lol..
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Supalemonhaze
Spore syringe hater.



Registered: 10/02/15
Posts: 6,725
Loc: 12" down Europe's butthole
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Re: 1st ever Agar Inoculation, Squirt or Swab? [Re: Mad Season]
#23194858 - 05/06/16 08:57 AM (8 years, 8 months ago) |
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I thought the long term water submersion would do the same job as forced hydration.
Never came across the charcoal but I see it can be found in health shops and shit, I might give it a go one day. Germinating spores on agar gives better peace of mind than fishing out mycelium from grain jars or LCs, that's for sure.
Thank god venom isn't required in this hobby, we got no snakes here .
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