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OfflinePBJ710
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Re: 1st Time working w/ Agar, Weak, Watery looking Myc from MS syringes? [Re: manlord]
    #27098509 - 12/19/20 10:39 PM (3 years, 1 month ago)

Quote:

manlord said:
Ok correct me if I'm wrong, but after streaking in a zigzag pattern, I'd want to watch for specific colonies to develop and then select strongest myc from multiple colonies? Cuz if I let it develop too much wouldn't the colonies like run into each other and maybe be harder to select healthy myc from the outside edges? Roughly how long after putting spores (MS or print) to agar do you typically take your first transfer? I know it varies, but. I've heard 1-2 days after you start seeing growth?




I try to get a transfer from a spore plate when the colony is about .75" and not overlapping other colonies - take a tiny sample from the leading edge.  It can take weeks to show any progress sometimes, but after growth starts showing 1-2 days is a good estimate.


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OfflineChardRich
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Re: 1st Time working w/ Agar, Weak, Watery looking Myc from MS syringes? [Re: PBJ710]
    #27099374 - 12/20/20 01:45 PM (3 years, 1 month ago)

Quote:

PBJ710 said:
Quote:

manlord said:
Ok correct me if I'm wrong, but after streaking in a zigzag pattern, I'd want to watch for specific colonies to develop and then select strongest myc from multiple colonies? Cuz if I let it develop too much wouldn't the colonies like run into each other and maybe be harder to select healthy myc from the outside edges? Roughly how long after putting spores (MS or print) to agar do you typically take your first transfer? I know it varies, but. I've heard 1-2 days after you start seeing growth?




I try to get a transfer from a spore plate when the colony is about .75" and not overlapping other colonies - take a tiny sample from the leading edge.  It can take weeks to show any progress sometimes, but after growth starts showing 1-2 days is a good estimate.




Lmao what :waitnonevermind:


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Offlinemanlord
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Re: 1st Time working w/ Agar, Weak, Watery looking Myc from MS syringes? [Re: PBJ710]
    #27101563 - 12/21/20 07:43 PM (3 years, 1 month ago)

Hey could you look at these plates and tell me what you think this is? I switched up my agar recipe and getting the same result from my B+ MS syringe as all the other agar plates I've tried with this one. Watery, milky looking growth that starts after just 1-2days but doesn't look like anything I'd want to transfer..
Is this just contaminated myc? Or is the growth itself 100% contam? any suggestions guys, these plates I put MS spores on about 36hrs ago. The PE plates I did at the same time show now growth yet, and EVERY B+ plate I've made has started growing SO fast, in this exact same way. Wondering what is up here:



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OfflinePBJ710
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Re: 1st Time working w/ Agar, Weak, Watery looking Myc from MS syringes? [Re: manlord]
    #27101798 - 12/21/20 10:24 PM (3 years, 1 month ago)

Milky looking growth on plates is usually bacteria especially if it shows up that quick - if there is any healthy growth, you should be able to transfer away from the bacteria, but it may take a few days or more to show up.  Would it be possible to get some better pics of it laying flat on a light colored surface?


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Offlinemanlord
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Re: 1st Time working w/ Agar, Weak, Watery looking Myc from MS syringes? [Re: PBJ710]
    #27101821 - 12/21/20 10:54 PM (3 years, 1 month ago)

I just removed the lids off the petris in front of the flowhood for a couple seconds and took 2 pictures, hopefully these ones are better.
  It could just be a bacteria ridden syringe, every single plate I've made (20+) have grown just like this, milky, and very quickly. I'd reallyyyy appreciate it if you'd watch this plate over the next 1-3 days with me just so
I can show you the growth and if there is ANYTHING healthy let me know and I'll transfer it asap. 
  Just wanna make sure I'm not crazy and that this syringe is in fact the problem.



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OfflinePBJ710
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Re: 1st Time working w/ Agar, Weak, Watery looking Myc from MS syringes? [Re: manlord]
    #27101934 - 12/22/20 02:25 AM (3 years, 1 month ago)

If they are growing that much in 36 hours, it's probably not mycelium. 

What agar recipe did you use?


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Offlinemanlord
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Re: 1st Time working w/ Agar, Weak, Watery looking Myc from MS syringes? [Re: PBJ710]
    #27102662 - 12/22/20 03:16 PM (3 years, 1 month ago)

10g agar,
10g Light malt extract
400ml water
Normally I use 500ml water, as well as 1g nutritional yeast. But I remade the agar with less water and no yeast to make sure it's the syringe and not my agar. Every single B+ plate has grown exactly as these 2.


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Offlinetiptrippy
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Re: 1st Time working w/ Agar, Weak, Watery looking Myc from MS syringes? [Re: manlord]
    #27102698 - 12/22/20 03:46 PM (3 years, 1 month ago)

Unfortunately that does not look like myc either. I suggest knocking up 1 BRF half pint jar with your syringes each. Let them get to 100% colonization and wait for invitro pins. Birth them in front of your flowhood and transfer the pins to fresh agar plates.

If your sterile tek and agar recipes are done properly you should at least be getting some sort of myc growth. What do your syringes look like? Can you see any black spores in them?

Is your flowhood built properly? Have you done the lighter test? Just shooting out ideas here.


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Re: 1st Time working w/ Agar, Weak, Watery looking Myc from MS syringes? [Re: manlord]
    #27102756 - 12/22/20 04:33 PM (3 years, 1 month ago)

Make more agar, it's great practice and getting clean spawn on agar is core in the hobby. Many of my first agar plates looked horrible like this, today I'm hustling out the shrooms like magic. You'll get there, hang in there and keep making transfers.

No fingers in the juice, boii


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Willpower is the one true virtue



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Offlinemanlord
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Re: 1st Time working w/ Agar, Weak, Watery looking Myc from MS syringes? [Re: smalltalk_canceled]
    #27102811 - 12/22/20 05:18 PM (3 years, 1 month ago)

What possible mistakes could I be making in my agar that could cause odd growth like that? Only thing I can think that's wrong with my agar or my sterile tek etc is that my agar has some condensation on the top few lids. The bottom half or so are great, but the top 1-3 out of 10 has a bit of condensation, which ends up dripping down on the agar.
  However, I've got test plates out in the area my other plates are in and I havent seen any odd growth on them, no contams nothing. I wrap all plates in parafilm before they leave the flowhood area. And I only open my pressure cooker directly in front of the flowhood, agar stuffed with poly and tinfoil on top.
  Anything with my tek you would suggest changing? I've poured so many plates and wasted them from this syringe cuz every single one looks like this. At this point its only got like 1cc left in it and its not nearly as thick with spores as my PE syringe, but I can still see some in there. Maybe its just bacterial, I will know once this PE starts to germinate, hopefully it's healthy and then I know my agar is ok etc. Ill post some pics of these PE plates


Edited by manlord (12/22/20 05:25 PM)


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Re: 1st Time working w/ Agar, Weak, Watery looking Myc from MS syringes? [Re: tiptrippy]
    #27102815 - 12/22/20 05:23 PM (3 years, 1 month ago)

The flowhood is a Bonzai flowhood from an online vendor and comes with a warranty etc. so I hope it works well, haven't tried the lighter test whats that? It def prevents a lighter from lighting within a couple ft of it when it's on. I have it in a separate room, and non of my "test plates" have shown signs of contam. Only once I put this MS syringe to them, I also retried with PE and they didn't start showing growth so suspiciously quickly like the b+ have been (every time on like 20+ plates). So I have more hope for the PE, its been 48hrs and I can just barely see some growth on the PE plates, im gonna post pics soon so you guys can hopefully take a look for me. If I can isolate anything healthy, I will, but would rather ask you guys first. I have some grain jars showing healthy looking growth rn and have a spore print on the way, if I haveee to I'll do BRF cakes, almost outta syringe now


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Re: 1st Time working w/ Agar, Weak, Watery looking Myc from MS syringes? [Re: manlord]
    #27102831 - 12/22/20 05:35 PM (3 years, 1 month ago)

You are working from syringes.
They are always garbage imo
Just a step towards clean agar
If you have a flowhood and a clue, it points towards the syringes.

Are all germination from the syringe like this


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Edited by smalltalk_canceled (12/22/20 05:35 PM)


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Offlinemanlord
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Re: 1st Time working w/ Agar, Weak, Watery looking Myc from MS syringes? [Re: smalltalk_canceled]
    #27102845 - 12/22/20 05:44 PM (3 years, 1 month ago)

Yeah, from this syringe, every plate germinates exactly like these. All milky and crazy and they take off way too fast. I should have kept my other syringes until I got a good agar plate to transfer from but I didn't really think about the possibility that the entire syringe is contamed af. Hopefully the PE will give me something to start on agar thats healthy, cuz all the other syringes I put to grain already, a few of which are looking like healthy myc growth so I could always g2g if they make it


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Re: 1st Time working w/ Agar, Weak, Watery looking Myc from MS syringes? [Re: manlord] * 1
    #27102867 - 12/22/20 05:57 PM (3 years, 1 month ago)

One trick you could try is to use the last drops of the syringe on straight pced 2,5h grains and try to isolate from the first myc growth, back to agar.
Work from that

Since this looks like bacteria, it may help. There's a long explanation why but I'd rather get more high


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Willpower is the one true virtue



Edited by smalltalk_canceled (12/22/20 05:59 PM)


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OfflinePBJ710
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Re: 1st Time working w/ Agar, Weak, Watery looking Myc from MS syringes? [Re: smalltalk_canceled]
    #27102940 - 12/22/20 06:46 PM (3 years, 1 month ago)

I second tiptrippy's recommendation of BRF cakes for less than ideal syringes - BRF is somewhat resistant to bacterial growth and can be a reliable way of getting clone material.


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Offlinemanlord
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Re: 1st Time working w/ Agar, Weak, Watery looking Myc from MS syringes? [Re: smalltalk_canceled]
    #27103432 - 12/23/20 02:39 AM (3 years, 1 month ago)

How long do I have to let grain colonize before selecting a piece to put to agar? I see myc growth on my jars now, but it's not really mature myc yet. There are grains that are all white and colonized though, should I maybe wait until I do the 1/3rd shake? Or grab it asap cuz it's healthy?


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Offlinetiptrippy
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Re: 1st Time working w/ Agar, Weak, Watery looking Myc from MS syringes? [Re: manlord]
    #27103440 - 12/23/20 02:57 AM (3 years, 1 month ago)

Quote:

manlord said:
How long do I have to let grain colonize before selecting a piece to put to agar? I see myc growth on my jars now, but it's not really mature myc yet. There are grains that are all white and colonized though, should I maybe wait until I do the 1/3rd shake? Or grab it asap cuz it's healthy?




You could grab a single grain as soon as it germinated and colonized 1 grain. But if you open a grain jar you take the chance of contaminating the whole jar.


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OfflineHappinessStan
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Re: 1st Time working w/ Agar, Weak, Watery looking Myc from MS syringes? [Re: manlord]
    #27103461 - 12/23/20 03:17 AM (3 years, 1 month ago)

I had the same issues as you with an MS syringe. I saw someone here say to try brf pucks and that worked for me, spores germinated in a week and I just pulled some mycelium off in my SAB and put that to agar plates to clean up.
My little pucks are now fully colonised and I'm waiting for pins to clone and hopefully a cap to make a print for future agar use.
MS syringes are always dirty and the bacteria will germinate befote the mycelium; go to brf in sterilised jars, it works.


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Offlinejcm4620
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Re: 1st Time working w/ Agar, Weak, Watery looking Myc from MS syringes? [Re: HappinessStan]
    #27103467 - 12/23/20 03:24 AM (3 years, 1 month ago)

i think that syringe is so fucked even goin to grains then to agar may not work. im sure someone here can send him a cube print?? id do it but im out and aint grown a cube in a year n a hlf. but what i will do is if someone wants to send him a cube print il send you a pan print


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Offlinetiptrippy
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Re: 1st Time working w/ Agar, Weak, Watery looking Myc from MS syringes? [Re: jcm4620]
    #27103809 - 12/23/20 09:39 AM (3 years, 1 month ago)

OP shoot me a PM I'll send you some goodies.


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