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OfflineJaRRn
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It just gets worse and worse *Updated*
    #3572747 - 01/02/05 09:27 PM (11 years, 11 months ago)

Hi all,

OK since I live internationally I have to order prints and make my own syringes. My first grow I had 70% success rate (did oven tek). The next grow I made a glove box. Following exactly what I did last time making the syringe, except using a glove box my failure rate was 100%! I left all my jars to see what would happen, some went black others green, some orange & red, and a lot mixed. I did like 20 Pf Jars & gladware wbs tubs. Just wasted!


*Step 1 - Put everything in there, pre-wiped with disinfectant/bleach.
*Step 2 - With lid slightly raised, spray like crazy then quickly close and seal.
*Step 3 - Wait 24hrs then using sterile knife? scrap spores into pre boiled water using brand new tips and syringes suck up to make 4 syringes per print.
*Step 4 -Lids on needles, which haven't touch anything only the air.
*Step 5 - Done - open, leave syringes for 24hrs before injecting.

**I know my problems lay in the syringe making as my mold/contam probs start exactly at the injection points. U can see the myc start growing with black or green shit coming out of it. I use a PC to sterilize pf jars.

I doubt it was a contaminated print as it was from a sponsor.

I'm thinking it has to do with the knife scraping, the air, or water.

For the water I use a Pyrex jar which is pc'd for 1.5hrs. Left to cool.

For the knife it is pc'd then Alcohol swabbed when put in GB. The GB gets a mega wipe down of course.

Air, I have nothing to help the air except the spray disinfectant.

I try to work as quickly as possible.

Everything was wiped down with disinfectants, bleach all the usual stuff. Should I consider throwing this box away and just doing the lazy way?

Or whats a good model(s) of hepa filter and how to utilize it as efficiently as possible? SunBeam Hepa I'm on the brink of giving up growing.
I've have ordered 3 more prints to give it one last whirl. If I can get 50% success I will continue.

Any links help ideas would be so greatly appreciated. I will start a grow log with the prints when they arrive so we can see how successful I will be this time. *:Fingerscrossed:*

If I cant get a contamination free environment and a successful grow happening I will gladly give free prints to those who helped.


I will have 4 Strains. Aussie (freebie from TLG). T.E.D, B+ and GT.(spores101). None of these sponsors where involved in that other print.


**** UPDATE ****

I made this.


It creates so much wind that I have a hole in the corner of the lid, the air just gushes out madly. (Blu Tac corner) Top left. Gloves try to push out aswell. I just used it, went well. I wiped everything down with bleach - I mean everything!, flamed, the scaping knife, wiped with swab. Made 5 syringes from one nice big dark print from TLG. I can see lots of clumps in all 5 turned out quite even.


Thanks all Peace.


Edited by JaRRn (01/09/05 09:11 PM)


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OfflineSuntzu
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Re: It just gets worse and worse [Re: JaRRn]
    #3574464 - 01/03/05 09:45 AM (11 years, 11 months ago)

Sounds like you have the 'materials' covered. . .it's not unheard of to receive a contaminated print once in awhile. A good thing to include in your testing is a jar that is not inoculated [negative control]. This will help show that it is likely the print that is to blame, though the fact that all the contamination originated from inoculation points indicates this as well. Good luck next time, it'll work!


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OfflineJaRRn
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Re: It just gets worse and worse [Re: Suntzu]
    #3575948 - 01/03/05 07:49 PM (11 years, 11 months ago)

Thanks for the reply.

Where is a good link on making a flowhood or a glove box.
Whats a good model of a hepa Filter I can buy.
What should I look for. Im so lost on this topic.
http://www.sunbeam.com.au/products/product_details.cfm?rec_id=46&sec_id=24 can only find this one. Australia has nothing ffs. lol

I want clean air.


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Invisiblewoodzmun
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Re: It just gets worse and worse [Re: JaRRn]
    #3576003 - 01/03/05 08:02 PM (11 years, 11 months ago)

It looks to me like you have a very good glovebox there to me JaRRn. If the contams started at your inoculation points and seeing how good you sterility is I would say it was the print. Before you do anything else I would try another print. Sorry I can't answer you question about the Hepa, as all I use is a glovebox too.


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OfflineJaRRn
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Re: It just gets worse and worse [Re: Suntzu]
    #3580909 - 01/04/05 08:15 PM (11 years, 10 months ago)

Quote:

Suntzu said:
A good thing to include in your testing is a jar that is not inoculated [negative control]. This will help show that it is likely the print that is to blame, though the fact that all the contamination originated from inoculation points indicates this as well.




Does/can the myc growing, speed up the contaminations, like bring it out ? If I have a control jar, & just say it was the jar that was contam'd, the contam would start a few days later and from any point.?


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OfflineSuntzu
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Re: It just gets worse and worse [Re: JaRRn]
    #3583375 - 01/05/05 10:54 AM (11 years, 10 months ago)

Actively growing mycelium shouldn't bring out contaminants that wouldn't otherwise grow. An untouched negative control that is 'contaminated' should show contam growth throughout, or near the top, but not specifically at inoculation points. Another negative control that may be helpful is a 'fake inoculated' negative control. For this one, you just clean the needle as usual, stick it in the jar, but don't expel any liquid. Theoretically, this might show if you have a bad needle or inoculating environment [separate issue from the spore solution itself].


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OfflineJaRRn
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Re: It just gets worse and worse [Re: Suntzu]
    #3585616 - 01/05/05 08:36 PM (11 years, 10 months ago)

Good Idea! Im going to nail this problem.


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Offlinekiss
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Re: It just gets worse and worse [Re: JaRRn]
    #3590980 - 01/06/05 11:19 PM (11 years, 10 months ago)


Hey Jarrn,
Don't give up man! You'll nail it down.
My box is pretty much like yours, and i'm not worried about contamination getting a foothold. I've prep'd everything just like you...no worries here! Not yet anyway.
Sounds like the print you got was funky. This I can only imagine would have to be better then using the oven. maybe only a fraction...i don't know?
Maybe I'm wrong on this but, if ya stop to think about it an oven isn't creating new air, it's just taking the same air(surounding air) from the bottom and pushing it up so nothing falls in, or on the surface. However there are small currents that you can't see just swirling about...ain't no telling where it's gonna end up...my opinion anyway :crazy2:
I just think your chances are better in that there box :smile:
Keep pluggin at it


--------------------
see dick drink
see dick drive
see dick die
don't be a dick!


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OfflineJaRRn
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Re: It just gets worse and worse [Re: kiss]
    #3591009 - 01/06/05 11:26 PM (11 years, 10 months ago)

Yeah I really dont want to chuck me box as it took ages to build. I had 70% success first time, and you'd think it would be 100% with the GB. 0% success instead. Ive got that hepa (linked) coming today. Try again this weekend. Got an Aussie print to use, from TLG. Grown aussies before. Cant wait till I get my T.E.D, B+ & GT from 101. Aussies are boring me now lol. Thats why i can afford a trial with them.

Gotta get it right before I Use me new prints. I'd liked to do a grow log to but not one where everything contams lol.


btw I like your box. 5 for you.


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Offlinekiss
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Re: It just gets worse and worse [Re: JaRRn]
    #3592629 - 01/07/05 07:54 AM (11 years, 10 months ago)

Hey thx Jarrn,

Here's another pic. of it :smile:
I noticed you used tape to keep your gloves in place. I was gonna go with that, but I opted for zip-ties. I know what you mean about the construction of these things. Took me all day to build this little thing  :smirk:
Cutting the plexiglass top was a real trick, but all came out well.
Keep me updated on how you do. Maybe by the time you start (deppending  :rolleyes: ) I could try and send ya a print. I have nothing at the moment though...still trying to germinate.


--------------------
see dick drink
see dick drive
see dick die
don't be a dick!


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OfflineJaRRn
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Re: It just gets worse and worse *Updated* [Re: JaRRn]
    #3605119 - 01/09/05 09:16 PM (11 years, 10 months ago)

I mean Bump. Updated.


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Offlinenewton
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Re: It just gets worse and worse *Updated* [Re: JaRRn]
    #3639480 - 01/17/05 04:47 PM (11 years, 10 months ago)

jarren- question about your glove box... Looks nice BTW...

your hepa is blowing air INTO the box thru it's front vent?

you did say that the air is pumping out thru a hole in the top of the glove box...  so im assuming its all good.

all the hepas i've ever seen suck in the front and out the top/back,
guess i should go look at more hepas....  :smile:


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OfflineJaRRn
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Re: It just gets worse and worse *Updated* [Re: newton]
    #3640254 - 01/17/05 07:44 PM (11 years, 10 months ago)

I have the hole at the top and opposite side of the way the air flow is. I run it on turbo mode and the pressue it creates is huge. The Hepa is actually laying on its back. I wiped the glove box down with bleach, every square inch then run it while I prepare everything else. So The air has been running for around 15 mins. Check My grow log in my sig. So far things look good.


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OfflineJaRRn
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Re: It just gets worse and worse *Updated* [Re: JaRRn]
    #3655627 - 01/20/05 10:08 PM (11 years, 10 months ago)

I did say It just gets worse and worse didnt I !!!!!!!!!!!

Check my grow log...


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Re: It just gets worse and worse *Updated* [Re: JaRRn]
    #3677732 - 01/25/05 02:40 PM (11 years, 10 months ago)

Is that cotton you have in the GB holes ?
If so, you should change it. Don't get me wrong, the design and idea of your GB is good, but that stuff shouldn't be there. Once opened, any contam spore can attach to that cottony surface and you'll not be able to wipe it out when cleaning.

MAIA


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Spiritual being, living a human experience ... The Shroomery Mandala



Use, do not abuse; neither abstinence nor excess ever renders man happy.
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