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OfflinePeteyboy
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First time with agar....Ps Subs
    #23486444 - 07/28/16 04:33 PM (7 years, 6 months ago)

Hey guys, It's my first time with agar, this all occurred 12 days ago.... I used pasty's no pour tek with green food coloring. I used a Ps sub syringe to knock up the dish in an SAB. But, I accidentally shot like .5 cc into the dish....newb mistake...but, I can see quite a bit of cloudy white growth but not over the whole surface just about 70%, I can also see rhizo growth at the edge of the agar going up the plastic...

so I have a couple questions...
1. Was it bad to use that much solution?
2. The white fuzz I see is not swelling in the center like the pics I see on the site, but does appear to be mycellium, not sure why?
3. Should I wait for the whole surface to be colonized before I take a wedge?
4.what would you suggest would be my next best step for Ps. Subs?

Sorry for all the questions, just totally new at this, and couldn't find a lot about Subs on agar on the site.

Also couldn't get clear picture cause of plastic...


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Invisible2shoes
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Re: First time with agar....Ps Subs [Re: Peteyboy]
    #23486678 - 07/28/16 05:41 PM (7 years, 6 months ago)

I would say make a transfer and grow that out. Keep your first plate in case you need to go back. Pics even shitty ones can help.


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Re: First time with agar....Ps Subs [Re: 2shoes]
    #23486793 - 07/28/16 06:11 PM (7 years, 6 months ago)

Your next step would be to transfer some clean wedges to moist cardboard, then woodchips, then spawn to an outdoor woodchip bed.


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OfflinePeteyboy
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Re: First time with agar....Ps Subs [Re: mindbentempire]
    #23487018 - 07/28/16 07:05 PM (7 years, 6 months ago)

Quote:

2shoes said:
I would say make a transfer and grow that out. Keep your first plate in case you need to go back. Pics even shitty ones can help.




OK sounds like a solid plan, thanks alot man! I'll see if I can get a somewhat decent pic. I wanna pop the top off in my SAB to snap a pic but I didn't want to take any unnecessary chances with contamination. If I'm gonna take a wedge, that gives me good reason!!

Quote:

mindbentempire said:
Your next step would be to transfer some clean wedges to moist cardboard, then woodchips, then spawn to an outdoor woodchip bed.




So I can go straight to cardboard from agar? That sounds easier then going to grains, should I boil it or just get it real moist?


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Offlinemindbentempire
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Re: First time with agar....Ps Subs [Re: Peteyboy]
    #23487958 - 07/29/16 12:40 AM (7 years, 5 months ago)

You could go to grains too.
Up to you.
There's lots of attempted sub's grows on here that you can find. Just read and compare to what everyone has done.


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OfflinePeteyboy
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Re: First time with agar....Ps Subs [Re: mindbentempire]
    #23488377 - 07/29/16 06:36 AM (7 years, 5 months ago)

Yeah I been reading my ass off lol, I think my eyes have crossed!
My fav so far is olives thread on subs which was incredible I must say! But he's an Aussie so he has access to the coveted stem butts lol.

Does this sounds right to you...I mean cardboard seems like the way to go, it's simple and I think I read it's contam resistant.
Grains seem more vulnerable to contamination but provide more inoculation points when mixed with the wood substrate, providing faster colonization?

I guess I could put a wedge on cardboard and grains and compare the two!


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Re: First time with agar....Ps Subs [Re: Peteyboy]
    #23489723 - 07/29/16 03:57 PM (7 years, 5 months ago)

I don't know whether you've come across this. Have a read if you haven't

https://www.shroomery.org/forums/showflat.php/Number/7478182#Post7478182

You could substitute fresh mushrooms with your colonised agar dish.
I would make a few transfers to have backups, just in case it fails.

Keep updating what worked for you. :smile:

Here's another way to do it:
https://www.shroomery.org/forums/showflat.php/Number/11774724



Edited by mindbentempire (07/29/16 04:05 PM)


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InvisiblebodhisattaMDiscordReddit
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Re: First time with agar....Ps Subs [Re: mindbentempire]
    #23489789 - 07/29/16 04:18 PM (7 years, 5 months ago)

Last link in my signature. You know what you're doing already but just some things to look at


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InvisibleBunya
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Re: First time with agar....Ps Subs [Re: bodhisatta]
    #23489818 - 07/29/16 04:36 PM (7 years, 5 months ago)

:cool:


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OfflinePeteyboy
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Re: First time with agar....Ps Subs [Re: Bunya]
    #23492045 - 07/30/16 09:51 AM (7 years, 5 months ago)

O.K. guys i got a picture of the dish last night. I took two pieces from the original plate. I took the pieces from the bottom of the dish where the white is the thickest, basically split it in half I dropped those two pieces onto the center of two new agar dishes.




So now the two pieces that i cut out and put into two new dishes, are obviously sticking up off of the new agar. Is that ok


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Offlinespacechildo
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Re: First time with agar....Ps Subs [Re: Peteyboy]
    #23492059 - 07/30/16 09:55 AM (7 years, 5 months ago)

Quote:

Peteyboy said:





looks like pure bacteria to me :sad:


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OfflinePeteyboy
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Re: First time with agar....Ps Subs [Re: mindbentempire]
    #23492086 - 07/30/16 10:01 AM (7 years, 5 months ago)

Quote:

mindbentempire said:
I don't know whether you've come across this. Have a read if you haven't

https://www.shroomery.org/forums/showflat.php/Number/7478182#Post7478182

You could substitute fresh mushrooms with your colonised agar dish.
I would make a few transfers to have backups, just in case it fails.

Keep updating what worked for you. :smile:

Here's another way to do it:
https://www.shroomery.org/forums/showflat.php/Number/11774724





Mindbent, dude thank you! I hadn't seen any of those posts yet! The second one is a winner!! Talk about making it as easy as possible! I'm going to be using this laundry basket tek for sure. I highly doubt i will have the spawn ready in time for this year, But i will have the entire winter to multiply the woodchips, hopefully if all goes well by next fall i will have enough for a monster outdoor bed.


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OfflinePeteyboy
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Re: First time with agar....Ps Subs [Re: spacechildo]
    #23492101 - 07/30/16 10:06 AM (7 years, 5 months ago)

Quote:

spacechildo said:
Quote:

Peteyboy said:





looks like pure bacteria to me :sad:




Ahhh man...I sure hope not!
I accidentally squirted out between .5 cc and 1 cc onto the plate with a syringe. So there was a bit of solution rolling around on top of the plate. Which I was hoping is why the white is spread all over like that. But when you look at the side of the plastic where the agar meets the edge. You can see little rhizo arms reaching through and across the agar. I did not take a pic of that before i cut the pieces of agar out and transferred them.


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Invisiblemupetmower
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Re: First time with agar....Ps Subs [Re: Peteyboy]
    #23492204 - 07/30/16 10:33 AM (7 years, 5 months ago)

it is.. that plates i trash.

and you only need one drop. not 1 cc. try again.

or use a loop. flame loop, put 1 drop on loop to cool, one more drop on loop to load spores, touch loop to plate. everything in SAB, except flame.


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InvisiblebodhisattaMDiscordReddit
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Re: First time with agar....Ps Subs [Re: mupetmower]
    #23492220 - 07/30/16 10:40 AM (7 years, 5 months ago)



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OfflinePeteyboy
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Re: First time with agar....Ps Subs [Re: bodhisatta]
    #23492265 - 07/30/16 10:56 AM (7 years, 5 months ago)

:baseballfuck:


hmmm well i used an SAB, sterile procedures, and a sponsors syringe. Additionaly, I used the same techniques to knock up 30 jars, and have had zero contams in those jars. I know that contamination happens to the best of the best no matter how much precaution is used. So do you think the bacteria came from having too much spore solution on the agar? Or from sterilization techniques?

Hoping to refine my methods so as to prevent this from happening again...


Edited by Peteyboy (07/30/16 11:01 AM)


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InvisiblebodhisattaMDiscordReddit
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Re: First time with agar....Ps Subs [Re: Peteyboy]
    #23492297 - 07/30/16 11:11 AM (7 years, 5 months ago)

Spore syringes are not 100% clean. The more you use the higher the chance of a rogue spore or bacterium getting in. On agar almost everything shows up. In a jar of spawn you get a bit more leeway and on brf cakes even more


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OfflinePeteyboy
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Re: First time with agar....Ps Subs [Re: bodhisatta]
    #23492415 - 07/30/16 12:09 PM (7 years, 5 months ago)

Quote:

Trusted Cultivator said:
Spore syringes are not 100% clean. The more you use the higher the chance of a rogue spore or bacterium getting in. On agar almost everything shows up. In a jar of spawn you get a bit more leeway and on brf cakes even more





ahhh ok, well that makes a lot of sense, you are the man! I guess ill have to chalk the plates up to a learning experience. I know you outlined the technique above on how to use a syringe with agar plates, thank you for that. But is it safe to say that for sterility concerns prints are better to use with agar then syringes? Or is the risk the same?


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Re: First time with agar....Ps Subs [Re: Peteyboy]
    #23492423 - 07/30/16 12:13 PM (7 years, 5 months ago)

I like prints more. But they're both fine to use.

You can make a home.made inoculation loop or buy 500 disposable ones for a few dollars on ebay


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Re: First time with agar....Ps Subs [Re: bodhisatta]
    #23492461 - 07/30/16 12:25 PM (7 years, 5 months ago)

I just tossed my agar plates and wil try again using your method.

I'll make the homemade loop and look for a TEK...

buuuut now I'm worried even more...I was reading that thread you sent me and i came across something about enzymes in the jar. Most of my jars have what appears to be clear water sweating from the mycellium...some of the jars have accumulated a good amount. the thread said that t could be from too high temps and or bacteria :sad:...The temps did get into the high80s a few times over the past couple weeks. The ones that are sweating are either completely colonized or between 80 and 95% complete. I thought the sweating was normal so i dint think twice. Are the cakes doomed as well?


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Edited by Peteyboy (07/30/16 12:42 PM)


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Re: First time with agar....Ps Subs [Re: Peteyboy]
    #23492464 - 07/30/16 12:26 PM (7 years, 5 months ago)

Pictures?


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OfflinePeteyboy
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Re: First time with agar....Ps Subs [Re: bodhisatta]
    #23492665 - 07/30/16 01:46 PM (7 years, 5 months ago)



this is the jar with the most of the "water". It is also the first to completely colonize.


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Re: First time with agar....Ps Subs [Re: Peteyboy]
    #23492815 - 07/30/16 02:48 PM (7 years, 5 months ago)

I started three agar plates with 1cc each (galindoi), had the same worry about too much liquid on top, however two weeks later they were fully populated and looking good.  (I'll know better next time)
I transferred those to a batch of grain jars which are happily colonizing right now.  Keeping fingers crossed, by growth is looking good so far.


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OfflinePeteyboy
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Re: First time with agar....Ps Subs [Re: jasen]
    #23492887 - 07/30/16 03:28 PM (7 years, 5 months ago)

Quote:

jasen said:
I started three agar plates with 1cc each (galindoi), had the same worry about too much liquid on top, however two weeks later they were fully populated and looking good.  (I'll know better next time)
I transferred those to a batch of grain jars which are happily colonizing right now.  Keeping fingers crossed, by growth is looking good so far.




Did they look like the photo in this thread?


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Offlinemindbentempire
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Re: First time with agar....Ps Subs [Re: Peteyboy]
    #23492967 - 07/30/16 04:04 PM (7 years, 5 months ago)

Quote:

Peteyboy said:


this is the jar with the most of the "water". It is also the first to completely colonize.




Your jar looks good man. :thumbup:


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Offlinejasen
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Re: First time with agar....Ps Subs [Re: Peteyboy]
    #23492996 - 07/30/16 04:23 PM (7 years, 5 months ago)

Quote:

Peteyboy said:

Did they look like the photo in this thread?




Not exactly.  That picture looked a bit more... slimey?  After 4-5 days the extra liquid in mine either evaporated or was absorbed by the agar.  The mycelium that grew was more fuzzy/cottoncandy looking.

I didn't get any pictures of that stage, but this is one of my jars a few days after adding the agar slice...



Edited by jasen (07/30/16 04:39 PM)


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OfflinePeteyboy
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Re: First time with agar....Ps Subs [Re: mindbentempire]
    #23493391 - 07/30/16 06:34 PM (7 years, 5 months ago)

Quote:

mindbentempire said:
Quote:

Peteyboy said:


this is the jar with the most of the "water". It is also the first to completely colonize.




Your jar looks good man. :thumbup:




:bearbreakdance: Awesome man...thank you!

Also I'm working on jars now to do the laundry basket tek you so kindly introduced me too, Thank you again sir!


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OfflinePeteyboy
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Re: First time with agar....Ps Subs [Re: jasen]
    #23493401 - 07/30/16 06:38 PM (7 years, 5 months ago)

Quote:

jasen said:
Quote:

Peteyboy said:

Did they look like the photo in this thread?




Not exactly.  That picture looked a bit more... slimey?  After 4-5 days the extra liquid in mine either evaporated or was absorbed by the agar.  The mycelium that grew was more fuzzy/cottoncandy looking.

I didn't get any pictures of that stage, but this is one of my jars a few days after adding the agar slice...






Yeah my plate looked nothing like cotton candy lol...back to square one and  cooking up some agar! I'll get it right this time around!


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