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 One of Us
Stranger
Registered: 03/12/12
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 Re: New to agar  [Re: mushboy]
#26370541 - 12/09/19 09:32 AM (4 years, 4 months ago) |
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My thoughts exactly! Thanks so much!
I am amazed that I only got one good bit of healthy mycelium out of 10 plates that I swabbed (the other 5 not pictured have zero growth, mycelial or otherwise), and technically the only plate that worked for me, I did not swab/streak at all. Should I try to streak with a little more pressure next time?
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 Caps McGee
Grandaddy Smurfshack
Registered: 10/28/17
Posts: 14,357
Loc: ally known as ... 
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I was around 4oclock plate 5... first 2 look like mold second 2 look weak/meshed, and the left side of 5 is fucky: need to bounce quick like, before it sporelates...
Its not that surprising: pe is notoriously difficult to get clean culture from... moisture, bacteria, contaminant spores; all picked up with your "sterile" swab... softer agar(less powder, I suggest starting at 7g/500ml) will have more moisture present and often aids in germination: in fact, the 5 plates you dismissed may be your best looking yet... have some patience with them as spores CAN take up to 6 weeks to germinate
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TeaforTwo
Flight Attendant
Registered: 09/22/19
Posts: 1,005
Loc: Psychonaut City, Mile Hi!
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had some stuff on plates looked like those wispy ones, noob eyes thought i saw something different, transfered it and both turned pretty gross grey afterwards. that plate 5 good myc looks mighty fine, as mushboy and Caps said dont waste any time transfering to get away from that bad left side asap.
haven't worked with swabs, they seem weird to me lol. 
do you hydrate them somehow so as to streak plate or just trying to get some spores or tissue material to fall to plate and germinate?
I often see swab tip just cut off onto plate, like #5, do they work otherwise? how many plates might you get from a single well spored swab?
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Caps McGee
Grandaddy Smurfshack
Registered: 10/28/17
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Spores to transfer to the plate... some guys just snip the tip off directly onto a dish... some work solely with swabs, I personally only use them when I've recieved them (typically PE varieties in trade) or in situations like my first RW giveaway for the F9 generation: I only had one F9 print, so took 30 swabs and spread the love... it's just something else to keep up with imo, so I dont keep them on hand, only wrap and sterilize what I need, if and when I need it... one dark swab could potentially inoculate dozens of plates, but each streak represents a new vector for contamination to all subsequent streaks ... cant flame a swap between plates so have to be particularly thoughtful about your movement and handling
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Caps McGee
Grandaddy Smurfshack
Registered: 10/28/17
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AGAR ENVY! (Anything and All Things Agar!) [Re: Tormato]
#26370734 - 12/09/19 11:20 AM (4 years, 4 months ago) |
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Dont change the subject title plz, it makes it difficult for us to keep threads on track
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Caps McGee
Grandaddy Smurfshack
Registered: 10/28/17
Posts: 14,357
Loc: ally known as ...
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Re: AGAR ENVY! (Anything and All Things Agar!) [Re: Caps McGee]
#26370736 - 12/09/19 11:20 AM (4 years, 4 months ago) |
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3... muhbad
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A.k.a
Stranger
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Re: AGAR ENVY! (Anything and All Things Agar!) [Re: Caps McGee]
#26370790 - 12/09/19 11:52 AM (4 years, 4 months ago) |
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The fifth picture there looks like the early stages of the stuff I had just posted a plate of.
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 LAGM2020      
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TeaforTwo
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Registered: 09/22/19
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Loc: Psychonaut City, Mile Hi!
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Re: AGAR ENVY! (Anything and All Things Agar!) [Re: Caps McGee]
#26370793 - 12/09/19 11:55 AM (4 years, 4 months ago) |
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heh, was changed a week ago, you got me bit gunshy on that sorta mention so was going to fix it with my reply but forgot
need to quit fuckin off and get more plates poured, got some generosity coming our way and want to get some these exciting names on plates asap
time is now, going to pour 40 plates and make 2 dozen pasties. we have premix PDA and MEA, as well have LME and BRF with telephone agar-agar. what would you recommend breakdown as? (this type recipe to this many plates/pasties)
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Be well, friend. Stay Strong and Beautiful, Always, and in All Ways~
Look Forward, Seek Truth, Find Real, Go Further~
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Caps McGee
Grandaddy Smurfshack
Registered: 10/28/17
Posts: 14,357
Loc: ally known as ...
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Re: AGAR ENVY! (Anything and All Things Agar!) [Re: A.k.a] 1
#26370794 - 12/09/19 11:57 AM (4 years, 4 months ago) |
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Agreed... I'd expect it to explode in black specks any moment... might be a seasonal wave of that particular species or something... there's so much we dont understand about the dynamics of contamination... I even postulate one MIGHT be able to use a carefully timed introduction of bacteria to induce a stellar first flush: a sort of "one and done"...
My recipe doesn't change except for trade/sell plates... SAGARI gets 6g LME, and 4g agar powder/500ml cold water (telephone) ... trade/sell plates are standard MEA (10g/10g/500ml), bc it cuts down in moisture... simple, effective, limited inventory
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One of Us
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Re: AGAR ENVY! (Anything and All Things Agar!) [Re: Caps McGee]
#26370854 - 12/09/19 12:39 PM (4 years, 4 months ago) |
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Wow. Glad I got those transfers done. I took 4 small transfers from the 3-5 o'clock areas... unfortunately I didn't realize that a couple of plates I used that were pre poured from last week were contaminated already... one moldy, one bacterial.
The other two are fine, except one I forgot to cool the scalpel before taking the transfer (still new to this) but I'm sure it's ok regardless.
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Caps McGee
Grandaddy Smurfshack
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Posts: 14,357
Loc: ally known as ...
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Re: AGAR ENVY! (Anything and All Things Agar!) [Re: One of Us]
#26370930 - 12/09/19 01:19 PM (4 years, 4 months ago) |
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I go in hot, not an issue... do be sure to flame the blade from mid-sharp to point... I was having issues with bacteria until I realized I hadn't been completely sterilizing the center of the blade, and was going in sideways/using more of the blade than necessary to cut and transfer the wedge
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A.k.a
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Re: AGAR ENVY! (Anything and All Things Agar!) [Re: Caps McGee] 4
#26370999 - 12/09/19 01:58 PM (4 years, 4 months ago) |
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Finally decided to just commit.
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LAGM2020
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mushboy
modboy
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Loc: where?
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Re: AGAR ENVY! (Anything and All Things Agar!) [Re: A.k.a]
#26371006 - 12/09/19 02:01 PM (4 years, 4 months ago) |
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Feasoghorm
Registered: 10/24/18
Posts: 4,384
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 Re: AGAR ENVY! (Anything and All Things Agar!) [Re: A.k.a]
#26371205 - 12/09/19 03:44 PM (4 years, 4 months ago) |
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Quote:
A.k.a said:
Finally decided to just commit.
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dr_bizzarro
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Re: AGAR ENVY! (Anything and All Things Agar!) [Re: Feasoghorm]
#26371405 - 12/09/19 04:53 PM (4 years, 4 months ago) |
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Question:
I just did my first transfer on PDYA plates. I had cooked the PDYA up for the MS Syringe innoc round. I had twice as much PDYA as I needed, so I put it in a jar. Later, for the 1st transfer, I heated up the solidified PDYA, then PC'd it, and poured new plates.
My problem is, 3 days later the plates are still liquid. Not jelled. And I wonder if I'll get any growth at all, as I don't see any.
So, did I miss a memo somewhere that says you cannot cool then re-heat PDYA for plates? Would it have been better to just pour it all the first time, making empty plates then wrap them for later?
Or, am I worried for nothing? Just seems lie the agar should be solid, not liquid!
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trubblesome
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Re: AGAR ENVY! (Anything and All Things Agar!) [Re: dr_bizzarro]
#26371844 - 12/09/19 08:20 PM (4 years, 4 months ago) |
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couple i've got going right now:
cut from the same streak plate, from the same general colony. the top one had no visible growth to the naked eye but had at first glance, 75 or so cells on the surface at 40x. I had originally ID'd the small colony that had germinated nearby as having been from a relatively lower number of spores than some other spots on the dish (still a shitload, bottom pic), and thought it'd be neat to get this small slice further out as well. When I transferred it the slice was up higher in the dish on the new plate, so I could get in closer to it on the scope. As I focused further in to the transferred slice, I saw more and more strands, the 75 or so on the surface were actually hundreds, probably thousands. That was 5 days ago, this is where they're at now. There's a better candidate for something to grow out for a mono on a different plate, but I kind of want to chase this one some more.
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Bobbit
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Re: AGAR ENVY! (Anything and All Things Agar!) [Re: dr_bizzarro]
#26372941 - 12/10/19 11:17 AM (4 years, 4 months ago) |
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Quote:
dr_bizzarro said:
Question:
I just did my first transfer on PDYA plates. I had cooked the PDYA up for the MS Syringe innoc round. I had twice as much PDYA as I needed, so I put it in a jar. Later, for the 1st transfer, I heated up the solidified PDYA, then PC'd it, and poured new plates.
My problem is, 3 days later the plates are still liquid. Not jelled. And I wonder if I'll get any growth at all, as I don't see any.
So, did I miss a memo somewhere that says you cannot cool then re-heat PDYA for plates? Would it have been better to just pour it all the first time, making empty plates then wrap them for later?
Or, am I worried for nothing? Just seems lie the agar should be solid, not liquid!
Most in here pour the whole sleeve full of plates at once.
You CAN reheat (and re-PC) agar (tho yours sounds broken), and it works for many.
The problem I have with only pouring some plates from a sleeve is that the sterility of the remaining plates is compromised once open.
Just pour all your plates and wrap them all, and use as required. No need to refrigerate, though some do.
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meaculpaUIO
Registered: 08/26/19
Posts: 956
Last seen: 4 years, 4 months
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Re: AGAR ENVY! (Anything and All Things Agar!) [Re: Bobbit]
#26373058 - 12/10/19 12:44 PM (4 years, 4 months ago) |
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Esteemed colleagues,
Whats the best o'clock on my 'plate'?
Quote:
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Dabadoodledoo710
Dab Connoisseur
Registered: 03/04/17
Posts: 1,140
Loc: Zone 5b
Last seen: 27 days, 18 hours
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Re: AGAR ENVY! (Anything and All Things Agar!) [Re: meaculpaUIO]
#26373077 - 12/10/19 12:55 PM (4 years, 4 months ago) |
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Need a better picture than that. And is that agar nearly an inch thick?
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One of Us
Stranger
Registered: 03/12/12
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Re: AGAR ENVY! (Anything and All Things Agar!) [Re: Bobbit]
#26373082 - 12/10/19 12:58 PM (4 years, 4 months ago) |
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I have a quick question:
Is there any rush to transfer from a plate with a bacterial satellite colony? What I mean is, bacteria can only contaminate mycelium by contact, correct? Bacteria (or endospores) can't become airborne and contaminate an entire plate the way a sporulating mold can, right?
So, as long as the area you wish to transfer from is sufficiently far from the bacteria colony, it shouldn't be contaminated. Is this right?
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