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OfflineChicagoMan
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TC spore syringe or Agar slurry to knock up 20qt jars tomorrow..suggestions? Big PE and status pics!
    #23624203 - 09/07/16 10:56 PM (7 years, 5 months ago)

Hello shroomery!
Just started 2 66qt Mono's, 3rd time, last round used slurry BRF and although spawn ration was low I fruited about 3 dozen Giant PE some ranging from 80g up to my best yet of 111g.  They were huge but not many.

This time used 5qt wbs spawn in the 66's and in 24hr the two tubs are 30% colonized on top..looking very good

So question now, I'm soaking enough wbs for 20qt I plan on loading tomorrow.

I have two ways of inoculation since I foolishly didn't save any colonized grain...getting better, learning more every round..so please make suggestions!!!

Method 1: TC spore prints, make spore syringe, knock up 10 or all 20qt

Method 2: I have 3 clean Agar plates of a PE clone, looking great.  My thought was to take a distal sector of the Agar, put in 1/4 pint, add sterile h20, attach blender blades and "slurry" the colonized Agar wedge.  Suck slurry into syringe and inoculate 10qt with it

If I attempt method 2, I'll do 10 with spores 10 with agar/slurry syringe.

Sterility is going to make or break it obviously, but I'm big into Bleach, Lysol, glove, and at min a SAB, might make flow hood tomorrow if you all think it's that much better, and thus would improve chances considerably.

Please give any advise!!! I should have made a master and will this time but cannot have 19 sterilized qt wbs jars waiting on a master this round.

So they all need to be inoculated.

Any thought suggestions welcomed and appreciated
Thanks,
CM

Edited by ChicagoMan (09/08/16 12:33 AM)

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OfflineChicagoMan
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Re: TC spore syringe or Agar slurry to knock up 20qt jars tomorrow..suggestions? Big PE and status pics! [Re: ChicagoMan]
    #23624432 - 09/08/16 12:36 AM (7 years, 5 months ago)

Any idea why I had giant PE, almost 10 days fruiting, opposed to huge pinset? Slurry?

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Offlinephlanx
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Re: TC spore syringe or Agar slurry to knock up 20qt jars tomorrow..suggestions? Big PE and status pics! [Re: ChicagoMan]
    #23624460 - 09/08/16 01:03 AM (7 years, 5 months ago)

maybe you could put a chunk of clean colonized agar into each jar instead. If you have 3 clean plates, you should have enough to put a decent piece in each jar.

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OfflineChicagoMan
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Re: TC spore syringe or Agar slurry to knock up 20qt jars tomorrow..suggestions? Big PE and status pics! [Re: phlanx]
    #23624668 - 09/08/16 03:53 AM (7 years, 5 months ago)

If I do that there is only 1 inoculation point...shake at 10, 30, and 70%?

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OfflineChicagoMan
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Re: TC spore syringe or Agar slurry to knock up 20qt jars tomorrow..suggestions? Big PE and status pics! [Re: ChicagoMan]
    #23624688 - 09/08/16 04:05 AM (7 years, 5 months ago)

With a standard 90mm petri, circumference 283mm, I could get 20
2mm x 1.7??mm pieces, but is that going to be the most efficient?
10ml h2o plus a 1/6 sector of clean myc from petri, blended/"slurried" 1ml per jar ...1/4ml x 4 liquid innoculant locations per jar.... has to have far faster colonization times...or I am I crazy?

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OfflineChicagoMan
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Re: TC spore syringe or Agar slurry to knock up 20qt jars tomorrow..suggestions? Big PE and status pics! [Re: ChicagoMan]
    #23625113 - 09/08/16 08:49 AM (7 years, 5 months ago)

Is there  enough mycelium in a 1/6 of a dish, large outer section to inoculate 10 qt?

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InvisibleBunji Fungi
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Re: TC spore syringe or Agar slurry to knock up 20qt jars tomorrow..suggestions? Big PE and status pics! [Re: ChicagoMan]
    #23625143 - 09/08/16 09:04 AM (7 years, 5 months ago)

Make a liquid inocculant like you said, that will expand you plate way better. Also you should never use spores to grain, especially if you have clean plates


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Offlinemushpunx
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Re: TC spore syringe or Agar slurry to knock up 20qt jars tomorrow..suggestions? Big PE and status pics! [Re: ChicagoMan]
    #23625169 - 09/08/16 09:17 AM (7 years, 5 months ago)

If you have the option to use agar then donwant
t use spore syringe to grain.

The "agar slurry" you are describing is called Liquid Innoculant.  There are some really great Teks for that if you search the tern liquid innoculant.


inoculate your agar plates and after germination make transfers until they are clean enough to put to grain.
You could then transfer to a couple  plates and grow then out, then inoculate each quart with an agar wedge.



The best way to do it is to inoculate a couple grain master quarts with an agar wedge, let them colonize and then grain to grain transfer into the rest of the jars.


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OfflineChicagoMan
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Re: TC spore syringe or Agar slurry to knock up 20qt jars tomorrow..suggestions? Big PE and status pics! [Re: mushpunx]
    #23625325 - 09/08/16 10:24 AM (7 years, 5 months ago)

Thanks for the replies. I'm pretty familiar at this point with how ideally I should  expand my strains. Just haven't gotten completely there yet.
  However, didn't leave a jar as a grain master. (Should have)  And the Agar is the 2nd transfer, away from first plate.  I just didnt have any more plates when doing the transfers. (Buy more, I know)  Also why I only want to use 1 plate. 
As far as LC's  are concerned ,I have 2 going now, but they were only started a few days ago. LME pints, one from a tissue sample one from MS syringe.  Need to start 2 more I have waiting with some of the clean Agar.

Plate agar to LC is a tek I don't see much about, I've read a bit about taking a wedge and placing in the syringe body, but no additional water, "Just push hard" ...for one jar..not 10 to 20

Is it needed to pulverize the Agar with a blender or some method with a little water to get it into solution and in the syringe?

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OfflineChicagoMan
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Re: TC spore syringe or Agar slurry to knock up 20qt jars tomorrow..suggestions? Big PE and status pics! [Re: ChicagoMan]
    #23626599 - 09/08/16 05:51 PM (7 years, 5 months ago)

Can I PC the grain jars with a tyvek filter, then metal lid with holes in it....without the tin foil on top?  After injecting I'll cover with aluminum so the small holes in the tyvek have some coverage.

It's time to PC and wife won't be home for couple hours with foil....silly but thanks!

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InvisibleBunji Fungi
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Re: TC spore syringe or Agar slurry to knock up 20qt jars tomorrow..suggestions? Big PE and status pics! [Re: ChicagoMan]
    #23631432 - 09/10/16 06:25 AM (7 years, 5 months ago)

People don't talk much about LC because of the contamination risk, but if your agar is clean and you have good sterile technique it is a great innoculant. I've never heard of anyone putting agar in a dry syringe and I don't think that's going to work well, lol.

There is plenty of information on blenderless LI if you do a search. I do a tic tac toe drop with a couple extra cuts and dump it in sterile water and shake the shit out of it. It can be used pretty much right away so long as you don't use too much water.

And you will want foil on your jars to PC. You don't want to get the tyvek wet.


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Offlinemushpunx
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Re: TC spore syringe or Agar slurry to knock up 20qt jars tomorrow..suggestions? Big PE and status pics! [Re: ChicagoMan]
    #23634419 - 09/10/16 11:52 PM (7 years, 5 months ago)

Quote:

ChicagoMan said:
Can I PC the grain jars with a tyvek filter, then metal lid with holes in it....without the tin foil on top?  After injecting I'll cover with aluminum so the small holes in the tyvek have some coverage.

It's time to PC and wife won't be home for couple hours with foil....silly but thanks!




You definetly want tin foil covering your jars yea. It keeps excess moisture from getting in/filters dry



But NO, dont do that. Tin foil is no filter. You aren't supposed to inject thru a tyvek filter, you are supposed to crack the lid open and inject directly into the jar.
Holes in your lid let contams in

You have to use a still air box of course, which you should already be doing if you. Are working with grains.


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Offlinemushpunx
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Re: TC spore syringe or Agar slurry to knock up 20qt jars tomorrow..suggestions? Big PE and status pics! [Re: mushpunx]
    #23634440 - 09/11/16 12:03 AM (7 years, 5 months ago)

Blending up an agar wedge in sterile water (you need an Erberbach or similar home made.device) is called liquid innoculant.

Liquid culture is i nnoculated with agar and the liquid media gets colonized.


Really its best to inoculate your LC with a Liquid Innoculant. But if you cant a few small agar wedges will work.
There isn't much too it really you just have to be very, very clean.


I do my LC like this:

I blend an agar wedge in about 50ml of sterile water for 2 or 3 seconds in an Eberbach unit, which I pour into an Erlenmyer flask (with SFD under a hole in the cap or a big wad of polyfill as a filter) that contains the liquid broth.
I then place the LC on a magnetic stir plate and let it colonize for a few days.

My LC broth is just 10 grams of light malt extract in a litre of water sterilized at 15 PSI for 45 minutes


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