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Offlineiloveshitaki
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WBS not colonizing after 2 weeks
    #13995074 - 02/20/11 11:17 AM (12 years, 11 months ago)

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 :shrug:


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OfflineProfessorPinHead
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Re: WBS not colonizing after 2 weeks [Re: iloveshitaki]
    #13995177 - 02/20/11 11:44 AM (12 years, 11 months ago)


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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Offlineiloveshitaki
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Re: WBS not colonizing after 2 weeks [Re: ProfessorPinHead]
    #13995687 - 02/20/11 01:29 PM (12 years, 11 months ago)

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 :shrug:


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OfflineProfessorPinHead
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Re: WBS not colonizing after 2 weeks [Re: iloveshitaki]
    #13995844 - 02/20/11 02:07 PM (12 years, 11 months ago)

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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