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OfflineDabrit
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Consensus on how many CC's of MS solution per quart jar of WBS?
    #23836726 - 11/15/16 07:57 PM (7 years, 2 months ago)

Preferably looking for answers from trusted cultivators.

Reason I ask I was reading some guides before suggesting 1-2cc's of MS solution per quart jar and I had a fair number of jars with contamination. This time around I followed Franks 12 steps tek to the letter other than adding a little gypsum while soaking. so I only used 8 drops per jar.

It's been days since I inoculated and so far no signs of colonization??

B+ variety Cubes btw if that matters.


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Invisibleazur
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Re: Consensus on how many CC's of MS solution per quart jar of WBS? [Re: Dabrit]
    #23836732 - 11/15/16 07:59 PM (7 years, 2 months ago)

Zero.
Use agar instead


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OfflineDabrit
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Re: Consensus on how many CC's of MS solution per quart jar of WBS? [Re: azur]
    #23836742 - 11/15/16 08:02 PM (7 years, 2 months ago)

Quote:

azur said:
Zero.
Use agar instead




This was not helpful, I have syringes to use up first and a lot more study before starting with agar. I also do not own a laminar flow hood and am lacking funds to purchase one currently.


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Offlinetump
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Re: Consensus on how many CC's of MS solution per quart jar of WBS? [Re: azur]
    #23836751 - 11/15/16 08:06 PM (7 years, 2 months ago)

Im no tc but agar is the correct way to do things. And to answer your real question 1/10 ml of spore syringe is enough per quart. Ive divided it out and done so before.


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Re: Consensus on how many CC's of MS solution per quart jar of WBS? [Re: Dabrit]
    #23836754 - 11/15/16 08:06 PM (7 years, 2 months ago)

When ive done ms on grains I use 1/4 cc per quart.

You had high contam rate cause spores to grain is risky. Not cc overdose.

You dont need a flow hood for agar. Just a SAB which u should be using anyway


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Invisibleazur
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Re: Consensus on how many CC's of MS solution per quart jar of WBS? [Re: Dabrit]
    #23836757 - 11/15/16 08:06 PM (7 years, 2 months ago)

Ok. Well, keep 6th grading it and in a few months you'll look back and think about how helpful it could have been.
You can do agar work in a still air box. If you can make jello, you can do agar work.


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OfflineKenetic
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Re: Consensus on how many CC's of MS solution per quart jar of WBS? [Re: Dabrit]
    #23836761 - 11/15/16 08:09 PM (7 years, 2 months ago)

I'm gonna say..... one drop of spore solution to agar (or you can streak)

I did syringe to grain for a few years and while I grew shrooms, I also had unpredictable results and grew other things with it, mainly bacteria.  btw, You can pour agar and do all your work in an upside-down tote, commonly referred to as a sab.  I do this easily all the time now.

With that said, if all I had was grain jars and syringes I would go for 1-2 cc per jar.  Be sure to shake the hell out of that syringe first, and flame the needle in between jars.  Try to shoot the spores on the glass so you can see the growth when it appears and not freak out because all the myc is growing in the middle of the jar and you can't see it.


--------------------
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Re: Consensus on how many CC's of MS solution per quart jar of WBS? [Re: Dabrit]
    #23836762 - 11/15/16 08:09 PM (7 years, 2 months ago)

A SAB is all you need.

if your doing whole grain (not pf tek) then you will have trouble with many contams that are present in your syringe.

The prints that those syringes were made from were made in non sterile environments making them have all kinds of nasties.

BRF cakes are very resilient to those contams and since they are left in solid form and not broken up as spawn they will fruit pretty well when directly inoculated with a ms syringe. Not so With grain because it harbors bacteria better and will be broken up when spawned giving the contams a step up on your myc.

Agar is very easy in a SAB and will make much less work and failure for you in the end.

Check out the Pastyplate tek in the link in my sig for a good start on agar.


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Re: Consensus on how many CC's of MS solution per quart jar of WBS? [Re: azur]
    #23836767 - 11/15/16 08:10 PM (7 years, 2 months ago)

Quote:

azur said:
Zero.
Use agar instead




:whathesaid: agar is the best. However if you must use spores I would only use a drop or two. Less is more.


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OfflineDabrit
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Re: Consensus on how many CC's of MS solution per quart jar of WBS? [Re: tombosley8]
    #23836826 - 11/15/16 08:30 PM (7 years, 2 months ago)

Quote:

tombosley8 said:
A SAB is all you need.

if your doing whole grain (not pf tek) then you will have trouble with many contams that are present in your syringe.

The prints that those syringes were made from were made in non sterile environments making them have all kinds of nasties.

BRF cakes are very resilient to those contams and since they are left in solid form and not broken up as spawn they will fruit pretty well when directly inoculated with a ms syringe. Not so With grain because it harbors bacteria better and will be broken up when spawned giving the contams a step up on your myc.

Agar is very easy in a SAB and will make much less work and failure for you in the end.

Check out the Pastyplate tek in the link in my sig for a good start on agar.




Ok thanks for the helpful advice, however in order to agar I need to have fruits to clone from in the first place do I not? I do not have any harvests that I can clone to the agar?

I always use a glove box, and flame sterilize between inoculation's.

So if starting from scratch would I then need access to LC or do I put MS solution onto a petri dish with agar and then keep transferring the mycelium until I have a rhizomorphic sector and then transfer to a grain jar?


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InvisiblePastywhyteMDiscord
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Re: Consensus on how many CC's of MS solution per quart jar of WBS? [Re: Dabrit]
    #23836832 - 11/15/16 08:31 PM (7 years, 2 months ago)

Put a drop of ms solution on the plate and let it germinate.


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OfflineDabrit
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Re: Consensus on how many CC's of MS solution per quart jar of WBS? [Re: Pastywhyte]
    #23836849 - 11/15/16 08:36 PM (7 years, 2 months ago)

Quote:

Pastywhyte said:
Put a drop of ms solution on the plate and let it germinate.




Ok, In your experience how many myc transfers do you usually do to get a clean mono culture sector ready for grain transfer?

Obviously being a noob I'm also not fully familiar what to look for when it comes to contaminates on agar in the jars it's pretty simple if it's bright white ropey/ fluffy with no signs of a milky white/greyish look or obviously anything green/blue/orange/red etc than you're golden.

I'm pretty sure half of my last batch all had cobweb mold that had taken over about 40% of the contaminated jars.


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Re: Consensus on how many CC's of MS solution per quart jar of WBS? [Re: Dabrit]
    #23836875 - 11/15/16 08:46 PM (7 years, 2 months ago)

You don't need an isolate. You just need a clean culture. You can get a verified clean culture in one transfer sometimes.


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OfflineKenetic
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Re: Consensus on how many CC's of MS solution per quart jar of WBS? [Re: Dabrit]
    #23836876 - 11/15/16 08:46 PM (7 years, 2 months ago)

since you plan on doing ms to grain I wouldn't worry about obtaining a monoculture.  You need agar for that


--------------------
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Re: Consensus on how many CC's of MS solution per quart jar of WBS? [Re: Kenetic]
    #23836945 - 11/15/16 09:12 PM (7 years, 2 months ago)

I wouldn't worry about obtaining a monoculture even though I do use agar.

Your better off fruiting a ms culture with slightly narrowed genetics after a transfer or two or three to clean it up and then taking some clones from those grows to agar for isolating certain traits but even then you won't have a mono culture most of the time.
just very narrowed genetics.

This will give you consistent results.

First fruiting from Ms(on agar) is a much easier way of obtaining better cultures than trying to obtain monocultures from MS because it gives you a good idea of what you are isolating and gives you a lot of variety to choose from.


I am sure there are situations where monoculture could be amazing but for someone just getting the feel of things it is way over the top IMO.


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OfflineDabrit
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Re: Consensus on how many CC's of MS solution per quart jar of WBS? [Re: Pastywhyte]
    #23837436 - 11/16/16 12:20 AM (7 years, 2 months ago)

Quote:

Pastywhyte said:
You don't need an isolate. You just need a clean culture. You can get a verified clean culture in one transfer sometimes.




Ok thanks. Just to confirm if I go the agar route do I just put a drop of ms solution onto the agar or do I actually inject the agar?

And then once I see obvious mycelium growth with no other signs of contaminates in the sector of the petri dish do I let the whole sector grow out before transferring to a grain jar?


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OfflineDabrit
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Re: Consensus on how many CC's of MS solution per quart jar of WBS? [Re: azur]
    #23837445 - 11/16/16 12:24 AM (7 years, 2 months ago)

Quote:

azur said:
Ok. Well, keep 6th grading it and in a few months you'll look back and think about how helpful it could have been.
You can do agar work in a still air box. If you can make jello, you can do agar work.





Ok thanks for making me come to the realization it's time to move onto the next step in mycology.

To confirm I did get 7 successful jars with ms before but had 5 jars go bad with what looked like cob web mold. Also lost my last batch of Pf tek Amazonian ms syringe to it as well.

So by using just a drop of ms solution per sector of a petri dish of agar I stand a much better chance of remaining contaminate free gotcha.


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Offlinewtfcrazymofo
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Re: Consensus on how many CC's of MS solution per quart jar of WBS? [Re: Dabrit]
    #23837599 - 11/16/16 02:54 AM (7 years, 2 months ago)

Agar is easy as 123.


--------------------
If you want to eat->https://www.shroomery.org/forums/showflat.php/Number/8553541

Bag sealers are to bulky (my hood isn't that big)
https://www.shroomery.org/forums/showflat.php/Number/28622922




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InvisibleMad Season
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Re: Consensus on how many CC's of MS solution per quart jar of WBS? [Re: wtfcrazymofo]
    #23838394 - 11/16/16 10:45 AM (7 years, 2 months ago)

Agar should technically be the same difficulty as the pf tek. Just 3 ingredients, mixed together at a ratio, just like the pf tek. Malt extract agar is my preferred choice, and that's 2g malt extract, 2g agar, and 100 ml of water.


--------------------
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Re: Consensus on how many CC's of MS solution per quart jar of WBS? [Re: Mad Season]
    #23838422 - 11/16/16 10:58 AM (7 years, 2 months ago)

ditch the glove box and/or remove the gloves from the glove box and you have a SAB.
Spitball Jedi's SAB TEK

You should read this too!
Spiball Jedi's The Basics

Glove boxes make a ton of turbulence because of the piston effect. With six inch holes and no gloves attached there is very little turbulence when moving your hands in and out of the sab. Also you shouldn't be flame sterilizing inside the box as that creates currents from the heat blowing contams into your work.


Edited by tombosley8 (11/16/16 10:59 AM)


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OfflineDabrit
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Re: Consensus on how many CC's of MS solution per quart jar of WBS? [Re: tombosley8]
    #23838658 - 11/16/16 12:11 PM (7 years, 2 months ago)

Quote:

tombosley8 said:
ditch the glove box and/or remove the gloves from the glove box and you have a SAB.
Spitball Jedi's SAB TEK

You should read this too!
Spiball Jedi's The Basics

Glove boxes make a ton of turbulence because of the piston effect. With six inch holes and no gloves attached there is very little turbulence when moving your hands in and out of the sab. Also you shouldn't be flame sterilizing inside the box as that creates currents from the heat blowing contams into your work.





When I say glove box it probably holds the same meaning as SAB. I don't have permanently attached gloves covering the holes into the box I just have to wholes cut in the side of it. I then just put on disposable rubber gloves and Tyvek coveralls cut off at the elbow with an elastic around my wrists and then enter into the box( After dipping them in 70% alcohol). And yeah I only flame sterilize outside of the box and then immediately but the syringe into the box for sterilization putting a couple of drops of ms onto the floor of the box to cool it down before inoculating.


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OfflineDabrit
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Re: Consensus on how many CC's of MS solution per quart jar of WBS? [Re: Mad Season]
    #23838671 - 11/16/16 12:15 PM (7 years, 2 months ago)

Quote:

Mad Season said:
Agar should technically be the same difficulty as the pf tek. Just 3 ingredients, mixed together at a ratio, just like the pf tek. Malt extract agar is my preferred choice, and that's 2g malt extract, 2g agar, and 100 ml of water.




I'm taking it this is the ratio to make just one petri dish worth of agar?

Coolio. I know the mixing part seems pretty elementary it's just the inoculating part and being able to decipher what is a clean culture and contaminates once I get going.

But I guess that's what this site is for a tool to learn and progress as a mycologist.


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InvisibleMad Season
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Re: Consensus on how many CC's of MS solution per quart jar of WBS? [Re: Dabrit]
    #23838676 - 11/16/16 12:16 PM (7 years, 2 months ago)

Depends. That's more like 2-4 dishes. 60mm dishes use like 25 ml


--------------------
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Re: Consensus on how many CC's of MS solution per quart jar of WBS? [Re: Dabrit]
    #23839028 - 11/16/16 02:42 PM (7 years, 2 months ago)

Quote:

Dabrit said:
Quote:

tombosley8 said:

Spitball Jedi's SAB TEK

You should read this too!
Spiball Jedi's The Basics






When I say glove box it probably holds the same meaning as SAB. I don't have permanently attached gloves covering the holes into the box I just have to wholes cut in the side of it. I then just put on disposable rubber gloves and Tyvek coveralls cut off at the elbow with an elastic around my wrists and then enter into the box( After dipping them in 70% alcohol). And yeah I only flame sterilize outside of the box and then immediately but the syringe into the box for sterilization putting a couple of drops of ms onto the floor of the box to cool it down before inoculating.




Ok kool, sounds like you get the idea.

One more thing is make sure to wipe all jars and tools with iso and before getting to work mist down the inside box with soapy water being sure to saturate all walls ceiling and floor. Read the SAB tek I linked to and it explains it very well.


--------------------


Edited by tombosley8 (11/16/16 02:43 PM)


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OfflineDabrit
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Re: Consensus on how many CC's of MS solution per quart jar of WBS? [Re: tombosley8]
    #23839994 - 11/16/16 08:33 PM (7 years, 2 months ago)

Quote:

tombosley8 said:
Quote:

Dabrit said:
Quote:

tombosley8 said:

Spitball Jedi's SAB TEK

You should read this too!
Spiball Jedi's The Basics






When I say glove box it probably holds the same meaning as SAB. I don't have permanently attached gloves covering the holes into the box I just have to wholes cut in the side of it. I then just put on disposable rubber gloves and Tyvek coveralls cut off at the elbow with an elastic around my wrists and then enter into the box( After dipping them in 70% alcohol). And yeah I only flame sterilize outside of the box and then immediately but the syringe into the box for sterilization putting a couple of drops of ms onto the floor of the box to cool it down before inoculating.




Ok kool, sounds like you get the idea.

One more thing is make sure to wipe all jars and tools with iso and before getting to work mist down the inside box with soapy water being sure to saturate all walls ceiling and floor. Read the SAB tek I linked to and it explains it very well.




Yeah that's pretty much what I do. I clean the inside with iso,  I use a 1:10 bleach to water ratio though and mist the walls ceiling and floor after putting the jars inside. And mist in the air inside the box and let sit for 10 minutes.  I then iso my gloves and such and then remove the foil lids and then iso all sides of the jar and lids and then begin innoculating


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Re: Consensus on how many CC's of MS solution per quart jar of WBS? [Re: Dabrit]
    #23841270 - 11/17/16 10:57 AM (7 years, 2 months ago)

Nice vids on the subject

and I think the bleach is overkill... just irritates my lungs at best IMO


--------------------


Edited by tombosley8 (11/17/16 01:44 PM)


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OfflineGilfoil
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Re: Consensus on how many CC's of MS solution per quart jar of WBS? [Re: tombosley8]
    #25427467 - 08/31/18 11:32 PM (5 years, 4 months ago)

Hey everybody who contributed here. I found this incredibly helpful and just wanted to say thanks! bump


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Re: Consensus on how many CC's of MS solution per quart jar of WBS? [Re: Gilfoil]
    #25427722 - 09/01/18 03:29 AM (5 years, 4 months ago)

:doublefacepalm:


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