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Offlinemr_dude
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First Attempt at Agar and Grain
    #23658540 - 09/19/16 11:45 AM (7 years, 5 months ago)

hiya dudes! some of you might remember me from this pretty piece a few months back, and im fulfilling my obsession and finally making the plunge to agar!

ill keep updating as things unravel, right now i have 9 inocculated plates (b+ cubes syringe i made from my print) and i was wondering if -judging by the picture attached, it would be a suitable time to do some transfers to some new plates and get me some quality mycellium. what do u think?



Follow-up: I've been reading up on the agar threads posted here, particularly about strain isolation, and came up with a few questions..

what the hell is my goal supposed to be? according to the videos and from what i understand, i should keep doing transfers to eliminate sectoring and get a nice white non-fuzzy looking chunk of mycellium. is that it?

i dont plan to go through the 500 plates of petris trying to achieve strain isolation, im just trying to give myself an advantage through using good mycellium as opposed to inoculate brf cakes with a syringe and pray for the best. with that in mind, should i transfer to grain all the samples of mycellium that grew nicely on agar? meaning should i try to fruit mycellium from my 2nd and 3rd and 4th agar transfer?

another thing i read is that i should not transfer more than 5 times on the same agar recipe, something about the mycellium's need to a different diet after a few transfers?

Edited by mr_dude (10/06/16 02:20 PM)

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Offlinejesustripped
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Re: Agar, grains, h-poo [Re: mr_dude]
    #23660396 - 09/20/16 12:07 AM (7 years, 5 months ago)

Yes it's time to transfer.

You answered your next question.  Your goal, for now, is to make clean mycelium to use as inoculant for grain jars rather than inoculate jars with a syringe and hope for the best.

To try to answer your next question...yes?  Say you take three transfers from the plate in the pic, and they look decent. So you take three more from each of those plates, and then either discard them or leave them around to pin and clone. Then the nine plates you just made look great, so you use some to inoculate grain jars, and some to transfer some of the best look myc to more plates.  Repeat. And yes, as long as you switch up your agar medium every few transfers, this can go on for quite some time.

I personally started agar about 6-8 weeks ago. I put three drops of spore solution on three plates, and since then I've gone through three 25 foot packages of foil, which means I've wrapped and pc'd approximately 75 plates/jars. From three drops of spore solution. So far. And there's really no end in sight.  I hope I've been of some help.


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row row row your boat gently down the stream, merrily merrily merrily merrily life is but a dream

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Offlinemr_dude
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Re: Agar, grains, h-poo [Re: jesustripped]
    #23665673 - 09/21/16 04:47 PM (7 years, 5 months ago)

You have certainly been of help. I did a few transfers yesterday to a new batch of dishes, luckily i have close to 50 dishes ready for inoculation and planning to go through them all at this phase. I'll post my progress here, wish me luck :mushroom2:

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OfflinePeteyboy
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Re: Agar, grains, h-poo [Re: mr_dude]
    #23665896 - 09/21/16 06:13 PM (7 years, 5 months ago)

I have been dropping my old plates into grain jars if they are clean...might as well get some fruit while your learning to work with agar....right?
The first plates I ever swiped turned out clean, I just spawned those jars, should hopefully have fruits in a little over a week...ahhhh yeeaaaaa


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Offlinemr_dude
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Re: Agar, grains, h-poo [Re: Peteyboy]
    #23665904 - 09/21/16 06:17 PM (7 years, 5 months ago)

Cant argue there brother! of course it would be nice to have fruits while learning since you never know when u might stumble upon some massive fruiters. Problem is I havent gotten to the grain tek yet and im kind of intimidated by the entire process, let alone spawning to a bulk substrate which i plan to do (and consequently building a monotub)

so yeah.. eeeek

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OfflinePeteyboy
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Re: Agar, grains, h-poo [Re: mr_dude]
    #23665923 - 09/21/16 06:25 PM (7 years, 5 months ago)

Dude it's actually pretty easy prepping grains, and substrate. Watch RR's video on prepping rye berries, makes it real easy, I've done four batches now and seem to have nailed every one. Just follow the steps to a T.
  But... from a newb to a newb, don't worry bout hpoo and pastuerizing right now. Just go with CVG buckettek. You will get experience with your PC doin agar and grains and eventually when you have a nice sack of shrooms, you can start messing around with the more complicated stuff since if you lose a project it won't hurt so much since your sitting on a bunch, know what I'm sayin
Building a mono is as easy as 123..

I know it sounds ridiculous, but the teks these guys have come up with on here make the whole thing from start to finish as easy as 123 haha.
  The one thing that I've found most challenging is the SAB work. Things can get frustrating in there as you learn your way through the necessary processes.


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Edited by Peteyboy (09/21/16 06:30 PM)

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Offlinemr_dude
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Re: Agar, grains, h-poo [Re: Peteyboy]
    #23668843 - 09/22/16 05:42 PM (7 years, 5 months ago)

you make a good point my dude, i cant imagine failing after all that time because of some noob mistakes with hpoo and pasteurizing, so better to have a backup stash first, thanks!

a tiny note about SAB: now dont bite my head off, i know how much all of you are keen on working in the utmost sterile environment for agar, and i idolize RR just as much as you and am very aware that he goes the full nine yards (shower, sleeves, mouthwash, FLOWHOOD!!!) but check this out.. i did half of my agar pouring outside of the SAB, and all of my transfers also in my living room with closed windows and NO air currents. i do my best sterilizing the counter and equipment and putting on a mask and gloves and an alcohol burner, that is because i absolutely could not see shit while using a SAB. shit would slip out of my hand and id pour outside the plate even :grin: i realize im greatly reducing my chances of success, but lets call it my small experiment. the first plates colonized fine with no trich for 10 plates, the second batch are colonizing as we speak and still nothing. i wanna see just how much i can get away with being sloppy.

Edited by mr_dude (09/22/16 05:50 PM)

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OfflineBobabouy
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Re: Agar, grains, h-poo [Re: mr_dude]
    #23668868 - 09/22/16 05:52 PM (7 years, 5 months ago)

Chances are the plates made outside a SAB are not clean, and chances are you may never know it.. Until it's too late! Go to the big box store and buy some thin plexiglass or equivalent. Cut a large square out of your SAB and cover it with the clear plexi. Use an adhesive type caulk to secure it and problem solved, you can see! Also if stuff is slipping and sliding in your hands, wait till the alcohol has evaporated from your gloves before you handle things. Get the hang of it and you'll be much better off.

Edited by Bobabouy (09/22/16 05:53 PM)

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Offlinemr_dude
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Re: Agar, grains, h-poo [Re: Bobabouy]
    #23668903 - 09/22/16 06:03 PM (7 years, 5 months ago)

:frown:

deep down i always knew.. RIP
i should get started on a new and improved still-air-box then
wish me luck

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OfflineBobabouy
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Re: Agar, grains, h-poo [Re: mr_dude]
    #23668909 - 09/22/16 06:05 PM (7 years, 5 months ago)

Good luck. Yeah, if you miss bacteria in a plate and then you expand that plate with transfers, you could be having problems down the road and have no clue what is causing it.

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Offlinemr_dude
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Re: Agar, grains, h-poo [Re: Bobabouy]
    #23680184 - 09/26/16 03:13 PM (7 years, 5 months ago)

Im not pursuing this just rather curious. first i thought these plates (result of first transfer) were infested with cobweb since they appeared to be growing in 3d rather than 2d. they appeared to be bulging from the agar and not leveled. but now im not sure, do they look alright to u?




Edited by mr_dude (09/26/16 03:23 PM)

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Offlinemr_dude
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Re: Agar, grains, h-poo [Re: mr_dude]
    #23680236 - 09/26/16 03:24 PM (7 years, 5 months ago)

I realized they are not clear enough, so for the sake of science and since i should've tossed them already, here are a couple of more pictures with the lid off. notice the fuzziness? doesnt look myc.. or does it? in the second picture, i took a paper clip and just lightly scratched at it, and it all came "off"


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InvisibleBoogieman47
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Re: Agar, grains, h-poo [Re: mr_dude]
    #23680599 - 09/26/16 05:46 PM (7 years, 5 months ago)

You just wasted that plate that is agar and the myc will just come right off of it that was a good clean plate


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Offlinemr_dude
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Re: Agar, grains, h-poo [Re: Boogieman47]
    #23683113 - 09/27/16 12:39 PM (7 years, 5 months ago)

Quote:

noob47 said:
You just wasted that plate that is agar and the myc will just come right off of it that was a good clean plate




well i still have 4 more that look exactly the same. to tell u the truth i dont know anymore where to go from here. the dudes said they're all probably contaminated and i should toss them and start over, and now ur saying its a clean plate. should i keep going with my transfers in a glove box based on these plates?
im unclear about one thing: if a plate comes off looking like my plates pictures above, does that rule out the possibility of their having any contaminents? in other words, if i get myc like i did and no trich, arent i on the safe side so far?

Edited by mr_dude (09/27/16 12:40 PM)

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InvisibleBoogieman47
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Re: Agar, grains, h-poo [Re: mr_dude]
    #23683134 - 09/27/16 12:53 PM (7 years, 5 months ago)

If you made those outside of a sab im surprised there isnt any contams and yes if the other dont have bacteria a milky looking growth or green or black you should be good do more transfers in a sab and if on the second transfer still nothing go ahead and noc up some jars


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Offlinemr_dude
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Re: Agar, grains, h-poo [Re: Boogieman47]
    #23683144 - 09/27/16 12:57 PM (7 years, 5 months ago)

alright thats good news, ill make second transfers tonight and post the updates :rolleyes:

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Offlinemr_dude
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Re: Agar, grains, h-poo [Re: mr_dude]
    #23687369 - 09/28/16 04:08 PM (7 years, 5 months ago)

In the meantime, how about someone explain to me the deal with isolates and more importantly preserving myc from tubs that were ideal performers.
this scenario is still half way through execution but im just thinking ahead.

- I did my first inoculations using 3 different syringes made from the same print.
- Not all plates showed the same growth so i started (and still am) doing transfers to get cleaner better myc.
- Assuming i end up with 6 plates of strong clean rhizomorphic myc and noc up 6 master grain jars which would each be used as spawn for a batch of jars.
- I then would spawn to bulk to 6 tubs and later move to fruiting conditions.

Assuming that in the end 2 tubs go bad, 2 tubs do not fruit, 1 does fruit with less than satisfactory results, and 1 does great. My question at this point are:
1- Could that happen? using clean myc from one plate would fruit well while clean myc from another plate would not (knowing that they originated from the same print and possibly same syringe)
2- how can i at that point reproduce the process in order to get clones of the tub that preformed well? should i keep the agar plate that was used to noc up grains? keep one of the jars and not use it in the tub?
3- assuming tub from plate #1 turned out a good achiever, does that mean any isolates originating from that plate would be equal or better?
4- assuming tub from plate #2 did not fruit at all, does that mean all the isolates made using this plate would also be crap?

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InvisibleBoogieman47
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Re: Agar, grains, h-poo [Re: mr_dude]
    #23687407 - 09/28/16 04:16 PM (7 years, 5 months ago)

Youre over thinking haha if all jars are clean you should get mushies in all tubs given that you give them good conditions once you get fruits pick out a few i usually take first mature and some from good clusters take a tissue sample and eat each one you cloned at different times to test potency then you go from there an isolate is many transfers and may not be worth it after a few transfers let the plate grow out and do another transfer on agar and put the rest into grain then do a g2g to 5 jars and test three of those the others keep in a fridge if the ones you tested give you good resluts g2g into 20 more jars and make 6 more tubs


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Offlinemr_dude
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Re: Agar, grains, h-poo [Re: Boogieman47]
    #23687448 - 09/28/16 04:25 PM (7 years, 5 months ago)

Oh.. that certainly seems simple enough. thing is i was reading PussyFart's thread on 10 PE isolates and it freaked me out seeing how some of these seemingly perfect isolates did not fruit at all :eek: any takes on that?

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InvisibleBoogieman47
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Re: Agar, grains, h-poo [Re: mr_dude]
    #23687518 - 09/28/16 04:50 PM (7 years, 5 months ago)

Could of been a few things the spawn could of been bacterial i cant remember if they were 10 different isolates or drom the same one if they were different then that isolate was shit... that thread was pretty dope for sure the only tubs i had that stalled were from being too bacterial or the sub was too warm when i spawned to it and it fuckes it up or hit with trich..


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