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OfflineJeeashrooma
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Registered: 03/23/11
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Mycelium growing in dark
    #14362262 - 04/27/11 04:54 PM (12 years, 10 months ago)

Hi All,

I'm 10 days into Amazonian mycelium colonization, and it seems to be growing in a grey color rather than the white that my B+ grew in. You'll notice the dark halo around the whiter interior blob of myc. Is this to be expected with Amazonians?


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Offlinek00laid
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Re: Mycelium growing in dark [Re: Jeeashrooma]
    #14362268 - 04/27/11 04:55 PM (12 years, 10 months ago)

thats contaminated.


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OfflinePerun
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Re: Mycelium growing in dark [Re: k00laid]
    #14362290 - 04/27/11 04:58 PM (12 years, 10 months ago)

Quote:

k00laid said:
thats contaminated.




:whathesaid:


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Offlineguywiththegun
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Re: Mycelium growing in dark [Re: Perun]
    #14362295 - 04/27/11 05:00 PM (12 years, 10 months ago)

Trichoderma, my friend.

:frown:


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Invisibleaqh
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Re: Mycelium growing in dark [Re: Jeeashrooma]
    #14362305 - 04/27/11 05:01 PM (12 years, 10 months ago)

Quote:

Jeeashrooma said:
Hi All,

I'm 10 days into Amazonian mycelium colonization, and it seems to be growing in a grey color rather than the white that my B+ grew in. You'll notice the dark halo around the whiter interior blob of myc. Is this to be expected with Amazonians?






You've got trich.
Better luck next time.


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OfflineJeeashrooma
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Re: Mycelium growing in dark [Re: aqh]
    #14362392 - 04/27/11 05:13 PM (12 years, 10 months ago)

Ugh. Its the same with the other 23 jars too. Looks like its back to the inoculation board.

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Offlinek00laid
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Re: Mycelium growing in dark [Re: Jeeashrooma]
    #14362518 - 04/27/11 05:32 PM (12 years, 10 months ago)

what are you innoculating them with?


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OfflineJeeashrooma
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Re: Mycelium growing in dark [Re: k00laid]
    #14362567 - 04/27/11 05:42 PM (12 years, 10 months ago)

Syringes I got from Ralphster - I wiped the needles between every jar, wore a mask, the works. Looks like the Trich is growing from the spore insertion point in every jar too -  musta been something in the syringes is all I can think at this point.

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Offlinek00laid
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Re: Mycelium growing in dark [Re: Jeeashrooma]
    #14362578 - 04/27/11 05:45 PM (12 years, 10 months ago)

you dont wipe needles.

alcohol does not kill trichoderma spores (as you can very well see)

you flame sterilize the needle. nothing is cleaner than red hot metal d00d.

after you eliminate the obvious variables you can blame the spore syringe.


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Offlineguywiththegun
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Re: Mycelium growing in dark [Re: k00laid]
    #14362677 - 04/27/11 06:03 PM (12 years, 10 months ago)

Tell us more about your procedure, because where there's one hole there might be ten. Are you using a glovebox or other method of still-air isolation to keep infections away?

You're not flame sterilizing (which means any nasties on the inside of that needle are still in it). You might be able to get away with wiping between jars (I did) but not until you flame sterilize. Even then, better safe than sorry.


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OfflineJeeashrooma
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Re: Mycelium growing in dark [Re: guywiththegun]
    #14363162 - 04/27/11 07:40 PM (12 years, 10 months ago)

I wiped down the kitchen table with rubbing alcohol, and put out the myc jars.

Washed hands and put on latex gloves and a surgical mask.

Undid syringe, wiped it down with RA, and started inoculation.

Didn't flame sterilize between jars, but did wipe the needle each time with RA.

Stored jars in a 5 gallon bucket with lid - wiped down inside and out with RA. The trich is growing in all 24 jars from the points of inoculation, so the contamination clearly happened before or at the time of the first injection.  I had the syringe undone, wiped and was injecting the first jar in all of about 30 seconds, with gloves and mask on.

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Offlineveda_sticks
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Re: Mycelium growing in dark [Re: Jeeashrooma]
    #14363182 - 04/27/11 07:46 PM (12 years, 10 months ago)

Quote:

Jeeashrooma said:
I wiped down the kitchen table with rubbing alcohol, and put out the myc jars.

Washed hands and put on latex gloves and a surgical mask.

Undid syringe, wiped it down with RA, and started inoculation.

Didn't flame sterilize between jars, but did wipe the needle each time with RA.

Stored jars in a 5 gallon bucket with lid - wiped down inside and out with RA. The trich is growing in all 24 jars from the points of inoculation, so the contamination clearly happened before or at the time of the first injection.  I had the syringe undone, wiped and was injecting the first jar in all of about 30 seconds, with gloves and mask on.




you have 2 major fails here. Not flame sterilising your needle and not using a glovebox. infact 3 fails, kitchens are the worst place for sterile procedures.


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Invisibleawakendone
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Re: Mycelium growing in dark [Re: Jeeashrooma]
    #14363248 - 04/27/11 07:59 PM (12 years, 10 months ago)

your sterile procedure definitely needs to be stepped up!
regardless, i suspect the syringe was contaminated anyways.

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Offlinek00laid
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Re: Mycelium growing in dark [Re: awakendone]
    #14363268 - 04/27/11 08:02 PM (12 years, 10 months ago)

flame sterilizing needles is a must.

built a simple still air box out of a cardboard box and some plastic wrap.


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Offlineguywiththegun
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Re: Mycelium growing in dark [Re: k00laid]
    #14363645 - 04/27/11 09:15 PM (12 years, 10 months ago)

If you don't feel like using a cardboard box, a big-ass clear bag will work as well, as long as you're careful about keeping it sealed. Moving the bag around can suck air in, I'd do the cardboard box every time.

The first time I knocked up jars, I cleaned the shit out of my kitchen, and I did it over the open oven (which was bunk tek and if I'd researched it I would have known). Every jar got lipsticked, and out the door went the hundred bucks of mine and other peoples money that I'd spent on the kit (which was my second mistake, start with your own equipment. Looks like you got that covered though.) You're halfway there dude, so don't give up. Your mind is totally in the right place, you just need to work on sterile procedure.

Look at it this way, you've got the jars, and your fruiting chamber. Spend another twenty bucks on a syringe, build a decent-but-cheap glovebox, and you're in business. And keep us updated!


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Offlinesteelmonkey
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Re: Mycelium growing in dark [Re: guywiththegun]
    #14364423 - 04/27/11 11:22 PM (12 years, 10 months ago)

I agree you did a few things unsterile but i'm leaning towards a contaminated syringe...23 straight jars is alot to blame on not flaming a needle but of course thats just my opinion..you really do need a glove box it will turn out to be invaluable especially since your new to sterile procedures


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Offlineafrosheen
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Re: Mycelium growing in dark [Re: awakendone]
    #14364545 - 04/27/11 11:44 PM (12 years, 10 months ago)

Quote:

awakendone said:
your sterile procedure definitely needs to be stepped up!
regardless, i suspect the syringe was contaminated anyways.



:whathesaid:


Even with awful sterile technique, you should have gotten at least 1 out of 24 jars that succeeded. Contaminated syringes...it happens sometimes.


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OfflineCaptainAhab


Registered: 12/25/09
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Re: Mycelium growing in dark [Re: afrosheen]
    #14365110 - 04/28/11 03:33 AM (12 years, 10 months ago)



I'll make you a deal on a gigantic flow hood... You come get it, you can have it. No more contam problems. You can stuff bodies inside of it, too. Can't beat that


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Offlineguywiththegun
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Re: Mycelium growing in dark [Re: CaptainAhab]
    #14365155 - 04/28/11 04:09 AM (12 years, 10 months ago)

Quote:

CaptainAhab said:


I'll make you a deal on a gigantic flow hood... You come get it, you can have it. No more contam problems. You can stuff bodies inside of it, too. Can't beat that




Seriously?
Where is you is, Ahab? :lol:


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OfflineCaptainAhab


Registered: 12/25/09
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Re: Mycelium growing in dark [Re: guywiththegun]
    #14366742 - 04/28/11 12:37 PM (12 years, 10 months ago)

Quote:

guywiththegun said:
Quote:

CaptainAhab said:


I'll make you a deal on a gigantic flow hood... You come get it, you can have it. No more contam problems. You can stuff bodies inside of it, too. Can't beat that




Seriously?
Where is you is, Ahab? :lol:




I'm semi-kidding. I got a really good deal on it not long ago, around 100 bucks. I had to go pick it up, though. I moved that bitch by myself. It weighs around 300 pounds. Stainless steel. Huge motor. It's a monster. Down 'round Los Angeles.


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