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iloveshitaki
Apprentice


Registered: 02/08/11
Posts: 56
Last seen: 12 years, 3 months
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WBS not colonizing after 2 weeks
#13995074 - 02/20/11 11:17 AM (13 years, 13 days ago) |
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Ok so I did my research and seemed to do everything correctly but none of the jars I knocked up after 2 weeks are doing anything.
I made 8 jars all together:
3 were prepared with WBS 36 hour soak and rinse and PC for about 90 mins
3 were prepared with WBS 24 hour soak and verm at the bottom PC for about 90 mins
2 were made by pouring boiling water over WBS and letting soak for 24 hours then rinsed (1 with verm one without)
For lids I made a big hole in the center stuffed with Polly fill and tyvek filters.
I let all of the jars sit in the PC overnight.
I knocked em up using fool proof inoc method (flame sterilized, rubbing alcohol soaked in sponge and no air contact with the needle) I put a bandaid over the hole after inoc.
Some of the jars I made very tight after shacking some I did 1/4 a turn.
I shook some jars after inoc others I just shook after once cooled and let to rest overnight.
I am keeping them around 77-80 in a tit (maybe will just leave in a dark closet for my next batch)
So do you think the pollyfil and tyvek are overkill?
Should I loosen my jars more? Should I keep them in light instead of darkness?
Also do you guys think it may just be a bad syringe? (Purchased from spores101... thinking I should switch vendors next time)
Any insight would be very helpful!
Thanks
-------------------- Yes I spelled shiitake wrong yes I am to cheap to pay $10 to fix it
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ProfessorPinHead
Trapped in the Archives....




Registered: 07/09/10
Posts: 4,568
Loc: F=G*m_1*m_2/r^2
Last seen: 4 years, 7 months
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Re: WBS not colonizing after 2 weeks [Re: iloveshitaki]
#13995177 - 02/20/11 11:44 AM (13 years, 13 days ago) |
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For lids I made a big hole in the center stuffed with Polly fill and tyvek filters.
I would have made 2 smaller holes and pulled polyfil through one of them. Then I would have put a dab of silicone in the other one and let it set for 24 hours while my WBS soaked. The silocone hole can then be used as an injection port.
I knocked em up using fool proof inoc method (flame sterilized, rubbing alcohol soaked in sponge and no air contact with the needle) I put a bandaid over the hole after inoc.
I would have done the same in a glove box..... But I would have instead used a alcohol soaked cotton ball over the injection port on the jars... Then just before the injection I would have removed it.
Some of the jars I made very tight after shacking some I did 1/4 a turn. I shook some jars after inoc others I just shook after once cooled and let to rest overnight.
I hope that was an LC that you shook. You don't want to do that with spores. It separates them. You want them to be together so they can germinate.
I am keeping them around 77-80 in a tit (maybe will just leave in a dark closet for my next batch)
This is ok. I would try and stay closer to the mid 70's and if you a talking about Shiitake you'll need lower temps. to fruit them.
So do you think the pollyfil and tyvek are overkill?
Yes. You only need one or the other. I would recommend polyfil.
Should I loosen my jars more?
I don't understand what you are asking here. If you are referring to the lids ont he jar then no. Keep them tight.
Should I keep them in light instead of darkness?
I the future let there be light.
Also do you guys think it may just be a bad syringe? (Purchased from spores101... thinking I should switch vendors next time)
It is possible but not likely. I would write it off as researcher error. -------
Two weeks is a long time though. I would plan on starting over and following some of the above suggestions.
Edited by ProfessorPinHead (02/20/11 11:50 AM)
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iloveshitaki
Apprentice


Registered: 02/08/11
Posts: 56
Last seen: 12 years, 3 months
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Thanks for the reply to all my questions! As far as syringes I am using MC spores as this is my first time. I only shook a few of the jars and left a few alone (to experiment.)
I was asking about how to tight to make the jars after they have been inoced. I heard to leave a little loose for the PC then tighten to shake, let sit for 24 hours then tighten for inoc.
Can the jars be left in light all the time or should I give them a set amount of time a day?
Also if my moisture content is off can that be a reason why I'm not seeing any signs? I don't even see any contams so I'm a little confused...
Thanks!
-------------------- Yes I spelled shiitake wrong yes I am to cheap to pay $10 to fix it
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ProfessorPinHead
Trapped in the Archives....




Registered: 07/09/10
Posts: 4,568
Loc: F=G*m_1*m_2/r^2
Last seen: 4 years, 7 months
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Re: WBS not colonizing after 2 weeks [Re: iloveshitaki]
#13995844 - 02/20/11 02:07 PM (13 years, 13 days ago) |
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Quote:
iloveshitaki said: Thanks for the reply to all my questions! As far as syringes I am using MC spores as this is my first time. I only shook a few of the jars and left a few alone (to experiment.)
I was asking about how to tight to make the jars after they have been inoced. I heard to leave a little loose for the PC then tighten to shake, let sit for 24 hours then tighten for inoc.
Can the jars be left in light all the time or should I give them a set amount of time a day?
Also if my moisture content is off can that be a reason why I'm not seeing any signs? I don't even see any contams so I'm a little confused...
Thanks!
Oh, I see. No you did it right you should leave them a little loose and tighten them all when they come out of the PC.
You can let them receive a 12/12 light cycle of ambient lighting. At 100% increase the lighting to 6500K.
You you can try injecting a little sterile water but like I said you are better off just starting over and making sure to build a better lid and not shake your jars after you inoculate them.
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