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InvisibleBigPapaMushroom
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Registered: 04/05/20
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Can someone check my agar?
    #26627618 - 04/25/20 04:29 PM (3 years, 10 months ago)

Hello All,

I am doing my first grow and I am curious what I have on my hands here. I did 12 pasty plates, and only one has growth that looks white enough to be mycelium. The rest have sort a green tint. Most plates also have a very viscous liquid of the same color as the growth. Could this be left over from the spore syringe? I did ~7 drops instead of 1.

Any help is appreciated, thanks!

P.S. Sorry the pictures are crap -- couldn't get my DSLR to focus, if anyone has advice on taking better pictures I am happy to repost better ones


Here is a picture of what I hope is mycelium:



And here is a picture of the liquid:

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OfflineGrowtech
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Re: Can someone check my agar? [Re: BigPapaMushroom]
    #26627686 - 04/25/20 05:01 PM (3 years, 10 months ago)

Well first of all, 7 drops is WAY too much, but maybe you already know this.  How long has it been since you spore juiced them?  I've seen it take up to 30 days to see germination even with fresh syringes.  4-10 days is about average, but sometimes it just takes awhile.


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Mycolo.gyGenetics & yield tracking system

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InvisibleBigPapaMushroom
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Re: Can someone check my agar? [Re: Growtech]
    #26627797 - 04/25/20 05:53 PM (3 years, 10 months ago)

Thanks for your reply. It's good to know it can take that long. I am heating my grow room to ~77°F in hopes that will help things go quickly.

I did 5 plates on 4/16 and these 12 on 4/18. I was actually mistaken, these plates have 3-4 drops; the first 5 had 7 drops. Im guessing this is still too much, what issues should I expect the excess spore solutions to cause?

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InvisibleRoger Clemency
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Re: Can someone check my agar? [Re: BigPapaMushroom]
    #26627943 - 04/25/20 07:05 PM (3 years, 10 months ago)

It looks like bacteria in that first pic. Hard to tell for sure though. THats the problem with spore syringes, you get some bacteria in the water and then you basically have every spore enveloped with contaminant so it’s hard to get the myc to grow out over the edge of bacteria, allowing you to grab a clean piece to xfer.


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Heaven starts right where hell ends

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InvisibleBigPapaMushroom
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Re: Can someone check my agar? [Re: Roger Clemency]
    #26628635 - 04/26/20 01:55 AM (3 years, 10 months ago)

So you think it's probably from the spore syringe? Should I just get a spore print and use an inoculation loop in the future?

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OfflineOvoidhunter
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Re: Can someone check my agar? [Re: BigPapaMushroom]
    #26628647 - 04/26/20 02:11 AM (3 years, 10 months ago)

You just used to much spore solution one drop would have been plenty enough. Really tho if you are working with agar it could be arguably easier to start from a spore print. Not that you can't use a syringe.


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Offlinepoisoned
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Re: Can someone check my agar? [Re: Ovoidhunter]
    #26628774 - 04/26/20 03:45 AM (3 years, 10 months ago)

meh, starting from a syringe isn't hard. But use only one drop. Use inoculation loop to spread it around and if you're too lazy for that, I used to just tilt the dish a few times to get that drop around a bit.


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How I do glass dishes

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OfflineFrankRhizo
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Re: Can someone check my agar? [Re: poisoned]
    #26628888 - 04/26/20 05:12 AM (3 years, 10 months ago)

:whathesaid:

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OfflinePiggyPig
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Re: Can someone check my agar? [Re: FrankRhizo]
    #26628927 - 04/26/20 05:54 AM (3 years, 10 months ago)

What I like to do when I'm trying to get things started quickly is to prepare a grain jar and squirt half a syringe of spore solution into the jar. Usually I will see mycelium starting within a week or so. Then I shake the jar until I see a colonised grain on top. Sterilize my tweezers, quickly open the jar and remove the colonised grain. Place the grain in the centre of an agar plate and it should grow out in a week to ten days. Using this method, I can have a plate ready for transferring within 2-3 weeks.

What I like to do is to remove the most aggressive piece of mycelium to transfer to another plate to clean up the culture. With every transfer, your culture will improve. Use the rest of the plate to noc up 6 grain jars with an equal size agar wedge in each. This method will allow you to always be improving your cultures while keeping other steps of the process moving along as well.

Spore syringe to agar is more common than the method listed above and should be attempted at the same time. When you're starting out, I find it best to keep starting new cultures every week. That way when something turns green or won't grow, you have something coming up right behind it. Nothing worse that losing a couple months work and having to start over.

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InvisibleBigPapaMushroom
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Re: Can someone check my agar? [Re: BigPapaMushroom]
    #26629821 - 04/26/20 12:58 PM (3 years, 10 months ago)

So now I am kind of confused. I woke up this morning and checked my plates to find a small, bright, white patch in most. Could the original growth be monokaryotic mycelium? I don't really get what that is supposed to look like. It doesn't make sense that mycelium could start growing in the middle of bacteria, but I clearly still have a lot to learn.

Again, sorry for the shitty picture.


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InvisibleBigPapaMushroom
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Re: Can someone check my agar? [Re: PiggyPig]
    #26629836 - 04/26/20 01:04 PM (3 years, 10 months ago)

I actually did inoculate two of my oat jars straight from the syringe, so it's good to know I can get some plates that way.

I used more spore solution than recommended because I figured it would be better to use it up rather than keep trying to store it, and I thought more spores would be better not worse. Lesson learned  :grin:

In true noob fashion I already fucked my pressure cooker up, so I gotta get another one. This has been quite a learning process. Not a bad way to quarantine.

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OfflineSockadin
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Re: Can someone check my agar? [Re: BigPapaMushroom]
    #26629839 - 04/26/20 01:05 PM (3 years, 10 months ago)

Have you looked at the picture? It is to blurry to tell what is going on.

How did you fuck up your pressure cooker?

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InvisibleBigPapaMushroom
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Re: Can someone check my agar? [Re: Sockadin]
    #26630008 - 04/26/20 02:10 PM (3 years, 10 months ago)

Fair enough mate. I have been having a hell of a time trying to get any of my cameras to focus through the curved side of a mini round. Here are all the pictures I have, maybe more will give a better idea:















I know they are shit quality, that is why I keep apologizing.

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OfflineOvoidhunter
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Re: Can someone check my agar? [Re: BigPapaMushroom] * 1
    #26630012 - 04/26/20 02:13 PM (3 years, 10 months ago)

Even with the quality of the pics I can tell you that it is bacteria and you should start over. Next time make way more plates to hedge failure. Don't forget to properly PC either. Good luck.


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InvisibleBigPapaMushroom
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Re: Can someone check my agar? [Re: Ovoidhunter]
    #26630093 - 04/26/20 03:03 PM (3 years, 10 months ago)

I appreciate the advice. Thank you for your time.

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OfflineGrowtech
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Re: Can someone check my agar? [Re: BigPapaMushroom]
    #26631981 - 04/27/20 11:23 AM (3 years, 10 months ago)

Well, spore syringes are inherently going to be contaminated (usually bacteria).  So even if your aseptic procedure introduced additional contaminants, I'm not sure "starting" over is going to solve it.  This is why you start on agar and do transfers until you get clean mycelium.  I'd say do that rather than start over.


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InvisibleBigPapaMushroom
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Re: Can someone check my agar? [Re: Ovoidhunter]
    #26688336 - 05/22/20 02:40 PM (3 years, 10 months ago)

Quote:

Ovoidhunter said:
Even with the quality of the pics I can tell you that it is bacteria and you should start over. Next time make way more plates to hedge failure. Don't forget to properly PC either. Good luck.










Hmm.... Same plates, ~30 days later. I'm glad you were so confident, but perhaps refrain from giving advice in the future if you can't actually see what was happening. (Jests aside, the curves of the mini round are inherently blurry -- it's impossible to take a clear picture of something that is visually distorted. So I don't really see this as either of our faults).

Anyway, breaking my PC turned out to be fortunate, otherwise I would have certainly started over. I am hopefully getting a new one soon (tomorrow?).

Can I transfer the plates without growth to grain jars? What should I do with the plates with growth?

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OfflineA.k.aM
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Re: Can someone check my agar? [Re: BigPapaMushroom] * 1
    #26688393 - 05/22/20 03:08 PM (3 years, 10 months ago)

Contaminated plates usually pin fast.

I’ve had a bunch grow over mold and pin on top of it and only in that spot.


Not saying yours is cuz I can’t tell from the pics but pins doesn’t mean clean.


I would pull the pin and put it on a new plate. Much better chance of clean myc that way.


--------------------
LAGM2020

Edited by A.k.a (05/22/20 03:11 PM)

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InvisibleBigPapaMushroom
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Re: Can someone check my agar? [Re: A.k.a]
    #26688580 - 05/22/20 04:07 PM (3 years, 10 months ago)

Well, it looks like I put my foot in my mouth on that one; I wasn't aware that mycelium could grow over mold.

Just for reference's sake, what is 'fast' pinning? I first saw pins about a 10 days ago, and they still fruited 3 days ago. pins appeared ~20-25 days after inoculation, probably.

I appreciate the advice, I'll give that a try. Just pop the lid and grab the pin with sterelized tweezers, then drop it on the new plate?

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