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OfflineAlCapone2k
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Agar plates - which one to inoculate with and which one to transfer from?
    #23818798 - 11/10/16 09:00 AM (7 years, 2 months ago)

Hey guys,

I have to different plates from the same generation (generation means number of transfers starting from an MS agar plate).

Both are the third generation, so tranferres three times.




The first plate (from up to down) is more hairy and not so white. The second one is fluffly and dense white.

Does that mean that the first one is rhizo and the second is tomentose?


But how would you go on?

1. Make another transfer with both of them?
2. Make Transfer from first one and inoculate with second one?
3. Make Transfer from second one and inoculate with first one?
4. Inoculate with both?



Regards,
Al Capone


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Re: Agar plates - which one to inoculate with and which one to transfer from? [Re: AlCapone2k]
    #23818817 - 11/10/16 09:06 AM (7 years, 2 months ago)

Whichever option 1-4 you like most, do that


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Re: Agar plates - which one to inoculate with and which one to transfer from? [Re: AlCapone2k]
    #23818820 - 11/10/16 09:07 AM (7 years, 2 months ago)

Tomatoes? Lmao jk, the top one looks a lot like the cobwebby looking contam I transferred away from yesterday :shrug:

Bottom looks good.


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OfflineAlCapone2k
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Re: Agar plates - which one to inoculate with and which one to transfer from? [Re: bodhisatta]
    #23818886 - 11/10/16 09:22 AM (7 years, 2 months ago)

Quote:

bodhisatta said:
Whichever option 1-4 you like most, do that





Thank you very much.

I thought it could be smarter to choose either the fluffy or the hairy one to inoculate with. But as both are ok, I'll try both :smile:

So the first/top one is also absolutely ok? :smile:




Quote:

Tomatoes? Lmao jk, the top one looks a lot like the cobwebby looking contam I transferred away from yesterday :shrug:

Bottom looks good.





Really? So you mean that the top one could have a contamination?


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Re: Agar plates - which one to inoculate with and which one to transfer from? [Re: AlCapone2k] * 1
    #23818942 - 11/10/16 09:35 AM (7 years, 2 months ago)

They're both tomentose and that's 100% ok
Rhizomorphic is not important at all
It will go rhizomorphic on grain anyway


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OfflineAlCapone2k
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Re: Agar plates - which one to inoculate with and which one to transfer from? [Re: bodhisatta]
    #23818963 - 11/10/16 09:40 AM (7 years, 2 months ago)

Yeah, I know, I read a lot of thread to this topic after using the search function earlier before opening this thread and you stated this many times.

I'll go for both.


Since I have you in this thread, just one question: I use your pour agar tek, boiling agar, filling into the kind of bottles you also use and then PCing. Let it sit in 50°C hot waterbath until the water bath itself cools down again to 50°C. Then I pour the plates.
Sometimes I get "hilly" plates, which means there is no plain agar in the plates but some small "hills" on it. It seems, that part of the agar inside the bottle is clumping. What am I doing wrong? Shall I shake the hell out of the bottle before pouring with it?


Regards.


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Re: Agar plates - which one to inoculate with and which one to transfer from? [Re: AlCapone2k]
    #23818987 - 11/10/16 09:45 AM (7 years, 2 months ago)

You may be having some agar get solidified while it's cooling down in the bottle. You have to swirl / mix the bottle now and then so that the temperature stays uniform otherwise youll get some agar solidified because it gets cold faster in some parts of the bottle. Or use enough water in the water bath to go over the level of agar in the bottle

You could try 52C too just a hair hotter


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OfflineAlCapone2k
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Re: Agar plates - which one to inoculate with and which one to transfer from? [Re: bodhisatta]
    #23818994 - 11/10/16 09:47 AM (7 years, 2 months ago)

Quote:

bodhisatta said:
You may be having some agar get solidified while it's cooling down in the bottle. You have to swirl / mix the bottle now and then so that the temperature stays uniform otherwise youll get some agar solidified because it gets cold faster in some parts of the bottle. Or use enough water in the water bath to go over the level of agar in the bottle

You could try 52C too just a hair hotter





Ah, ok. thank you very much! I'll try to swirl/mix it next time more :smile:

Regards


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Re: Agar plates - which one to inoculate with and which one to transfer from? [Re: AlCapone2k]
    #23819025 - 11/10/16 09:55 AM (7 years, 2 months ago)

Also if your water bath is half as tall as the agar in the bottle. The agar above the level of hot water may solidify


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OfflineAlCapone2k
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Re: Agar plates - which one to inoculate with and which one to transfer from? [Re: bodhisatta]
    #23819045 - 11/10/16 10:00 AM (7 years, 2 months ago)

Yeah, but if I remembered correctly, I always used a waterbath higher than the agar level in the bottle :smile:


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Re: Agar plates - which one to inoculate with and which one to transfer from? [Re: AlCapone2k]
    #23819203 - 11/10/16 10:40 AM (7 years, 2 months ago)

Quote:

AlCapone2k said:
Quote:

bodhisatta said:
Whichever option 1-4 you like most, do that





Thank you very much.

I thought it could be smarter to choose either the fluffy or the hairy one to inoculate with. But as both are ok, I'll try both :smile:

So the first/top one is also absolutely ok? :smile:




Quote:

Tomatoes? Lmao jk, the top one looks a lot like the cobwebby looking contam I transferred away from yesterday :shrug:

Bottom looks good.





Really? So you mean that the top one could have a contamination?



No im just saying I know mine was a contam because it popped up away from inoculation point. Im no pro just giving my input lol


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Re: Agar plates - which one to inoculate with and which one to transfer from? [Re: spirit_shadow]
    #23819292 - 11/10/16 11:08 AM (7 years, 2 months ago)

Interesting thread. I have a question or two.

This guy has two plates that look entirely different, and the vibe I'm getting is pull from wherever. Why is that? They sure do look different. Is that after 3 transfers and no obvious contams, should be good to use the whole plate even?

Thanks


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Re: Agar plates - which one to inoculate with and which one to transfer from? [Re: Moabfighter]
    #23819304 - 11/10/16 11:11 AM (7 years, 2 months ago)

If you're inoculating jars you use all the mycelium you can


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Re: Agar plates - which one to inoculate with and which one to transfer from? [Re: bodhisatta]
    #23819402 - 11/10/16 11:33 AM (7 years, 2 months ago)

You don't use the cleanest, prettiest little spot of the plate per jar? Don't you get a mixed bag using all those different genetics on the plate? Does it even matter?


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Re: Agar plates - which one to inoculate with and which one to transfer from? [Re: Moabfighter]
    #23819416 - 11/10/16 11:36 AM (7 years, 2 months ago)

you say "per jar" well if you have "per jar" for instance 4 jars you would be using 1/4 of the growth on the dish per jar I would hope.

hows it much different than using spores, it's still MS but the plates are limiting the genetics so it's like "limited MS"



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Re: Agar plates - which one to inoculate with and which one to transfer from? [Re: bodhisatta]
    #23819436 - 11/10/16 11:40 AM (7 years, 2 months ago)

I'm just unsure of the methods with agar. Do people usually inoculate like that? I think it seems maybe most people make LI or LC with their agar? It doesn't seem I've seen a lot of inoculating jars with plates. Maybe use a whole plate to inoculate a master, then G2G?

Sorry, I'm just trying to get a really good understanding of everything that I do. I'm trying to excel.


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Re: Agar plates - which one to inoculate with and which one to transfer from? [Re: Moabfighter]
    #23819451 - 11/10/16 11:42 AM (7 years, 2 months ago)

most people just use their agar, fewer people it seems to me make LC or LI. you sure can go ahead and do LC or LI

you can do a whole plate to a master and g2g. that's just as fast a LC or LI

there's kind of a million and a half ways to do things the right way once you use the right techniques.


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Re: Agar plates - which one to inoculate with and which one to transfer from? [Re: bodhisatta]
    #23819454 - 11/10/16 11:43 AM (7 years, 2 months ago)

you take tiny transfers when transferring on agar,
and use large wedges (or several smaller wedges in each jar) when inoculating grains.


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Re: Agar plates - which one to inoculate with and which one to transfer from? [Re: Moabfighter]
    #23819464 - 11/10/16 11:44 AM (7 years, 2 months ago)

I've been inoculation jars with plates but it's tedious and takes forever.  I'm trying an li in a couple days because I see from other people that it's pretty fast.


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Re: Agar plates - which one to inoculate with and which one to transfer from? [Re: Kenetic]
    #23819490 - 11/10/16 11:48 AM (7 years, 2 months ago)


i don't find it tedious but do what works for you


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Re: Agar plates - which one to inoculate with and which one to transfer from? [Re: bodhisatta]
    #23819517 - 11/10/16 11:53 AM (7 years, 2 months ago)

Good information thanks everyone


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Re: Agar plates - which one to inoculate with and which one to transfer from? [Re: bodhisatta]
    #23819559 - 11/10/16 12:03 PM (7 years, 2 months ago)

Rather than go 1 plate to GM, and g2g to 10 jars. I have found it effective to go agar to Li, and directly to 10 jars. Same end result, just getting there faster.

Additionally, when dealing with spawn that is a little bacterial you don't have to worry about g2g weakening and contaming jars.

My first grow was agar to grain, tiger drop 1.0 & 2.0 (tic tac toe) taking one plate per jar. Second go around I used Li and 1 plate in 10 jars. Definitely will be using Li for my 3rd go too
:2cents:


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Re: Agar plates - which one to inoculate with and which one to transfer from? [Re: Moabfighter]
    #23819569 - 11/10/16 12:05 PM (7 years, 2 months ago)

I'd rather g2g but I've been having issues with bacterial spawn for Some reason.  Dropping plates isn't that bad but it is slow as hell for me. I need quick colonization apparently and Li seems like it would solve my problem.


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Re: Agar plates - which one to inoculate with and which one to transfer from? [Re: lukehighwalker710]
    #23819572 - 11/10/16 12:06 PM (7 years, 2 months ago)

You do the blenderless LI? Seems easy enough for a beginner... add to jar of sterile water (boil water, pour in jar, put lid on, wait to cool?) add agar plate, shake, pour into ten jars?


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Re: Agar plates - which one to inoculate with and which one to transfer from? [Re: Kenetic]
    #23819587 - 11/10/16 12:09 PM (7 years, 2 months ago)

Quote:

kenetic said:
I'd rather g2g but I've been having issues with bacterial spawn for Some reason.  Dropping plates isn't that bad but it is slow as hell for me. I need quick colonization apparently and Li seems like it would solve my problem.




prep your grains better/differently so they're dry on the outside, PC them for longer, get better at doing sterile work.
LI wont solve problems with bacteria etc, its just another step which helps you if you got your shit down but can fuck up more for you if you dont.


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Re: Agar plates - which one to inoculate with and which one to transfer from? [Re: Moabfighter]
    #23819623 - 11/10/16 12:18 PM (7 years, 2 months ago)

Quote:

Moabfighter said:
You do the blenderless LI? Seems easy enough for a beginner... add to jar of sterile water (boil water, pour in jar, put lid on, wait to cool?) add agar plate, shake, pour into ten jars?



Um. I definitely PCd my jar of water for Li for like 30-40 min. But yes blenderless and yep thats the basic idea.


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Re: Agar plates - which one to inoculate with and which one to transfer from? [Re: spacechildo]
    #23819678 - 11/10/16 12:34 PM (7 years, 2 months ago)

How about this for 2nd transfer.




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Re: Agar plates - which one to inoculate with and which one to transfer from? [Re: Edmunter]
    #23819688 - 11/10/16 12:36 PM (7 years, 2 months ago)

why do you take such large transfers? are most of them just agar or do you take a large chunk of myc? remember the leading edge is barely visible.


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Re: Agar plates - which one to inoculate with and which one to transfer from? [Re: spacechildo]
    #23819704 - 11/10/16 12:42 PM (7 years, 2 months ago)

I just wanted to get those hand like strains on top.  This is 2nd transfer from spore germination.  Ive never got this far so quick


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Re: Agar plates - which one to inoculate with and which one to transfer from? [Re: Edmunter]
    #23819992 - 11/10/16 02:31 PM (7 years, 2 months ago)

Li is amazing ime :trippinbawelz:


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Re: Agar plates - which one to inoculate with and which one to transfer from? [Re: spirit_shadow]
    #23819994 - 11/10/16 02:32 PM (7 years, 2 months ago)

Quote:

spirit_shadow said:
Li is amazing ime :trippinbawelz:





How you doing it man?


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Re: Agar plates - which one to inoculate with and which one to transfer from? [Re: Moabfighter]
    #23819998 - 11/10/16 02:34 PM (7 years, 2 months ago)

Well I use pastys so once I have a clean plate I just inject it with sterile water and use the sterilized needle to kinda scratch the surface of the myc and suck it back up.


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Re: Agar plates - which one to inoculate with and which one to transfer from? [Re: spirit_shadow]
    #23820014 - 11/10/16 02:43 PM (7 years, 2 months ago)

Do you sterilize the water within the syringe? Or do you sterilize it in the jar and pour it inside the pastyplate?


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Edited by AlCapone2k (11/10/16 02:43 PM)


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Re: Agar plates - which one to inoculate with and which one to transfer from? [Re: AlCapone2k]
    #23820121 - 11/10/16 03:28 PM (7 years, 2 months ago)

I sterilize the water in the syringe.

1. Bring pot of water to boil.

2.pull syringe full of water.

3. Squrit that into the SINK. NOT the pot with boiling water.

4. Repeat steps 2 and 3 about 4 times.

5. Let sit till cool.


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Edited by spirit_shadow (11/10/16 03:36 PM)


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Re: Agar plates - which one to inoculate with and which one to transfer from? [Re: spirit_shadow]
    #23820147 - 11/10/16 03:36 PM (7 years, 2 months ago)

No concerns about heat resistant microbes that are pushed back into the water from the outer edged of the pot?

IIRC that was one of the problems of self made spore syringes?!


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Re: Agar plates - which one to inoculate with and which one to transfer from? [Re: AlCapone2k]
    #23820203 - 11/10/16 03:52 PM (7 years, 2 months ago)

nah heat and humid air rises. away from your pot.


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Re: Agar plates - which one to inoculate with and which one to transfer from? [Re: bodhisatta]
    #23820332 - 11/10/16 04:44 PM (7 years, 2 months ago)

The first plate is not very healthy growth the second is healthy tomentose growth, I wouldn't use the first one at all and use the 2nd one to either make more transfers or put it to grain


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Re: Agar plates - which one to inoculate with and which one to transfer from? [Re: frankwhite85]
    #23820436 - 11/10/16 05:32 PM (7 years, 2 months ago)

So toss the first one? :smile:

Maybe you are right, since another transfer wouldn't change anything. Either there is a hidden contamination (what I don't think) or it's just bad genetics, hm?


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Re: Agar plates - which one to inoculate with and which one to transfer from? [Re: AlCapone2k]
    #23820478 - 11/10/16 05:43 PM (7 years, 2 months ago)

Do what you want :vibin:


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Re: Agar plates - which one to inoculate with and which one to transfer from? [Re: AlCapone2k]
    #23820485 - 11/10/16 05:44 PM (7 years, 2 months ago)

time to put your research skills to use capone! who to trust....?


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Re: Agar plates - which one to inoculate with and which one to transfer from? [Re: spacechildo]
    #23820587 - 11/10/16 06:16 PM (7 years, 2 months ago)

Sorry guys, just don't want to make anything wrong, since it's my first grow. And contradictory statements can confuse at this stage of noobishness :laugh:

But you are right spacechildo, I think I know who to trust ... especially bhodisatta was always a very good advisor and always helpful :smile:

So as the whole community of course.


--------------------
Since I am from Germany, my english is not perfect but I try my best :smile:




My small Agar to Oyster Open Air Grow Trek


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OfflineEdmunter
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Re: Agar plates - which one to inoculate with and which one to transfer from? [Re: spacechildo]
    #23821897 - 11/11/16 04:34 AM (7 years, 2 months ago)

Quote:

spacechildo said:
time to put your research skills to use capone! who to trust....?




Yep there is some shit advice on here recently. 

So who do you trust.  Someone with 17000 posts or someone with 50?


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OfflineAlCapone2k
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Re: Agar plates - which one to inoculate with and which one to transfer from? [Re: Edmunter]
    #23822074 - 11/11/16 06:37 AM (7 years, 2 months ago)

I know, I know :wink:

Greetings to spain edmunter (iirc)


--------------------
Since I am from Germany, my english is not perfect but I try my best :smile:




My small Agar to Oyster Open Air Grow Trek


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OfflineEdmunter
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Re: Agar plates - which one to inoculate with and which one to transfer from? [Re: AlCapone2k]
    #23822165 - 11/11/16 07:22 AM (7 years, 2 months ago)

Quote:

AlCapone2k said:
I know, I know :wink:

Greetings to spain edmunter (iirc)



:wave:


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