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oxohawkoxo
Myconaught


Registered: 10/03/07
Posts: 23
Last seen: 12 years, 29 days
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Grrr....
#7532889 - 10/18/07 08:05 PM (16 years, 3 months ago) |
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Alright... So my first run was 7 jars BRF, they all got contaminated. Trych. Green mold. So I tried a second run, 6 jars, new syringe, different medium, same sterilization procedures and ... NOTHING ... literally, not even contaminates. So a third run, 5 jars with a BRF/WBS combo medium (had to use up the last of the BRF), a third syringe, and paranoid sterility. Cleaned the desk, the floor, my hands, flamed and alcohol between each jar plus worked very quickly. Now this is the most contaminated run of them all, actually it kinda looks like I inoculated them with spores for this blue/green mold; not the slightest sign of any mycelium.
Now I didn't use a still air box (I will next time). All 3 syringes were from a Shroomery vendor. ... Is it really this hard? Though I must admit I used to use metal lids with just coffee filters back in the day with limited success. I'm now using tyvek under metal plus tape(though not micropore, I'll get some of that stuff too) Two holes, one for injection and another for FAE.
Has anyone else had this level of trouble getting some spawn started?
Any crazy suggestions? Other than the micropore tape, and a still air box? I think I'm going to make my own syringes as well. I'm not sure I trust vendors...
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Chi Ro
Jive Ass Turkey


Registered: 09/22/07
Posts: 211
Loc: Right hurr!
Last seen: 12 years, 5 months
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It's really not that hard. My first two jars got contaminated and I have had no contaminations since.
...are you using a Pressure Cooker? You didn't mention it, and that would be the problem if you're just knocking up jars that haven't been sterilized.
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CtK
Stranger
Registered: 10/01/07
Posts: 31
Last seen: 12 years, 3 months
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What do you mean, "Two holes, one for injection and another for FAE."
Are you leaving an open hole in your jars for FAE? I was under the impression colonizing jars didn't need FAE. If you have a completely open hole in all of your jars, it would probably explain the amount of contaminated jars you're producing.
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LayYouIn
Taurus



Registered: 09/28/06
Posts: 4,402
Loc: Organ
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Quote:
oxohawkoxo said: Any crazy suggestions? Other than the micropore tape, and a still air box? I think I'm going to make my own syringes as well. I'm not sure I trust vendors...
it's not the vendors.
where at in your jars do they start molding? is it in the spots you inoculated? if not, then you might not be sterilizing correctly.
what did you do with the jars that did nothing? ime, contaminates show up pretty fast(within 2-3 days). so, when i have no mold after the 4th day, i expect white mycelium and it's usually what i get.
Quote:
CtK said: What do you mean, "Two holes, one for injection and another for FAE."
Are you leaving an open hole in your jars for FAE? I was under the impression colonizing jars didn't need FAE. If you have a completely open hole in all of your jars, it would probably explain the amount of contaminated jars you're producing.
the verm layer should act as a filter. i want to take the lid off of a jar after inoculating just to see what happens...
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oxohawkoxo
Myconaught


Registered: 10/03/07
Posts: 23
Last seen: 12 years, 29 days
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Re: Grrr.... [Re: LayYouIn]
#7533621 - 10/18/07 11:10 PM (16 years, 3 months ago) |
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Hi Guys,
Thanks for the responses. To answer a few questions, yes I have a pressure cooker. I generally put my jars (quart jars) in for an hour before taking them out. One of the jars that was contaminated in my first run was bought from a vendor pre-sterilized. I flamed and alcohol wiped immediately before inoculating that jar though.
In regards to the FAE I don't leave any holes in the lids. I make two holes in the metal caps, put tyvek underneath, and inoculate though one of the holes, then tape over it. The second hole is just so that I have enough gas exchange.
The jars that start molding at the sites where I've inoculated, which leads me to believe it's the syringes, or my sterilization methods. My last round are so severely contaminated that it looks as though I inoculated with spores for the blue mold that's in there. Though the holes are only open to the air for a few moments before I tape them over again. Hence why I think I may be in severe need of a still air box.
I'm in an apartment building and I'm not sure my vents even have filters on them. If a sterilized still air box would make up for the filters in the vents I'll go with the box, otherwise I could always look into a couple of filters.
The jars that did nothing, they're still sitting in a case untouched. They still have no sign of mycelium or contaminate. What are the odds of anti-fungals in a birdseed? My sterilization procedures for those jars is the very same as used for my first run and similar but less strict than used for the third and worst round of jars.
That's where it stands. I think the best steps are to get a still air box for inoculation, and maybe modify my lids?
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The shroomy 1
Luminous beings surround me




Registered: 03/27/07
Posts: 5,543
Loc: The Aether
Last seen: 5 months, 5 days
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How funny, I always, always keep a "control" jar just for these reasons! I suggest everyone do the same. Don't inoculate one of the jars that have gone through the same procedures from start to finish. Hell, I have a blank syringe that gets pressure cooked and I use that to inoculate a jar. (I do that when I make my own syringes). The control jar is very important to me. Let's me know how sterile my procedures are, and makes isolating the contamination point so much easier. That's just me.
The shroomy 1
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AMU Q&A thread.
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RoachMan
Old Man




Registered: 03/06/05
Posts: 2,083
Loc: Midwest
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Try putting the tyvek between the lid and ring.
(Not under the lid as this can allow the filter to become wet which allows contaminants to move through the layer.)
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veda_sticks
Cultivator




Registered: 07/29/07
Posts: 14,191
Loc: UK
Last seen: 4 years, 25 days
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Re: Grrr.... [Re: RoachMan]
#7534053 - 10/19/07 03:49 AM (16 years, 3 months ago) |
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Make yourself a glove box, when i first started i didnt have a glove box, my first run i had 2 jars that didnt grow, then the next batch 3 out of 4 contam, then the next batch 1 out of 3. Then i made a glove box, 2 jars nicely colinising.
Most syringes come with the needle seperate, the syringe could have got contamed in the process of removing the the cap and isntalling the needle.
Make yourself a glovebox, and this will eliminate a possibility.
Mould in your jars means that they are probably getting contamed at inoc or incubating.
You could also try disinfecting your incubating area, i have mine in a nice warm cupboard, i give that a spray with beach everynow and then.
-------------------- PF TEK - writeup by EvilMushroom666 Lets Grow Mushrooms - RogerRabbit & RoadKills website with sample videos plus the full PF TEK video series. Alot of great information - BUY THE DVD Cakes can and will pin! - So you think cakes suck for pins. Your wrong Franks Simple Coir/Verm Tek Franks Proper Pasturisation Tek Franks Spawning To Bulk - Monotub Professor Pinheads RTV Injection Port Tek Foo Mans No Soak WBS Prep Tek
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ThePyschonaut52
Stranger


Registered: 04/24/07
Posts: 982
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from my experience, most contaminates enter the syringe when you put the needle on. You need a glove box.
-------------------- "In god we trust..." -I guess we're screwed.
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RoachMan
Old Man




Registered: 03/06/05
Posts: 2,083
Loc: Midwest
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Quote:
veda_sticks said: Make yourself a glovebox, and this will eliminate a possibility.
Eliminate a possibility, yes, however, I've done 100's of open air inoculations without a contamination.
(Not that I've never had a contaminant, but my only contaminants were back when I was a noobie and didn't pressure cook my grains long enough.)
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simplemachine
Manfly


Registered: 09/14/03
Posts: 1,981
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Quote:
The jars that start molding at the sites where I've inoculated, which leads me to believe it's the syringes
I think you are right on this one. Your syringes should definitely be made in a still air box. Inoculating in there isn't as important, because its easy to clean whats OUTSIDE the needle, but you have be make a clean syringe first.
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