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Invisibleshroomybgood
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Best way to isolate AA+?
    #14327952 - 04/21/11 02:21 PM (12 years, 9 months ago)

I've got a tub of Albino A+ fruiting and she's really taking off. I've heard that AA+ is not an abundant fruiter so I really want to isolate this strain? There's got to be 500+ pins in there and more coming up daily. What's the best way to reproduce this effect? LC, cloning, agar...? I've had other successful grows but none like this. Please help me keep it going. Thanks.


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Offlinem_jacobs
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Re: Best way to isolate AA+? [Re: shroomybgood]
    #14328170 - 04/21/11 03:07 PM (12 years, 9 months ago)

**What I would do.

  • Take a few mature fruits, and clone to agar.
  • Take multiple samples from different fruit if you have the agar.
  • Grow them out on agar and clean them up if necessary.
  • Then go Ager to small jar of WBS.  Once WBS is 100%, I would do GLC.
  • From GLC I would do full size Quart and then go G2G.


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Offlinesteelmonkey
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Re: Best way to isolate AA+? [Re: m_jacobs]
    #14328341 - 04/21/11 03:42 PM (12 years, 9 months ago)

Yeah..what m_jacobs said


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Invisibleshroomybgood
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Re: Best way to isolate AA+? [Re: steelmonkey]
    #14328400 - 04/21/11 03:49 PM (12 years, 9 months ago)

How long can I store the agar for? I need to isolate so that I can bring it back at a later date. I guess I should have stated that in the OP. I figure I have about 3 weeks till my present project is finished at which point I will be going out of town for about 3-6 months. I want to isolate this so that I can replicate this winter. Thanks.


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Offlinesteelmonkey
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Re: Best way to isolate AA+? [Re: shroomybgood]
    #14328436 - 04/21/11 03:56 PM (12 years, 9 months ago)

Your best bet is to make a culture slant they will keep for years in the fridge


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InvisibleBarakanaten
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Re: Best way to isolate AA+? [Re: steelmonkey]
    #14328452 - 04/21/11 03:59 PM (12 years, 9 months ago)

I would add to take multiple clones from pins/clusters.

However many plates you think you might need make extra in case a few get contams.

I would take a few extra dishes and toss in a few quart jars and then G2G.

Meanwhile you can work on your isolation technique.

Take a dish that is 1/2 or 3/4 colonized and take a piece
of tissue from the leading edge and transfer that to a new dish,
do that 3-4  times and keep the original dish and store it away for later.

grow those petris out on cakes or small treys
until you find what you want then remember where it came from and start isolating that...

here's a great vid:super:

just remember to label and keep track of what is what,
always make extra and make a master slant of the good stuff.


:sunny:


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OfflineRogerRabbitM
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Re: Best way to isolate AA+? [Re: shroomybgood]
    #14328512 - 04/21/11 04:11 PM (12 years, 9 months ago)

Quote:

shroomybgood said:
There's got to be 500+ pins in there and more coming up daily. What's the best way to reproduce this effect?




This is not necessarily a good thing.  If you have 500 pins, most will abort, and you may end up with less total product than if you were starting out with less, but more survivable pins.

Remember, the name on the print or syringe means little.  What matters is the exact pairing of compatible hyphae, and the best way to get there is via strain isolation on agar.  After isolating a dozen or more strains, fruit each one to determine the best performers.
RR


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Invisibleshroomybgood
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Re: Best way to isolate AA+? [Re: RogerRabbit]
    #14328609 - 04/21/11 04:28 PM (12 years, 9 months ago)

Thanks RR for ruining my vision!lol I just watched your isolation video and I think I'm going to start dabbling a bit. Will a GB be sufficient for doing the transfers? I have a small hepa in my room too. Also, is there anyway I can slow the pinning to get more out of my fruits?


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InvisibleBarakanaten
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Re: Best way to isolate AA+? [Re: shroomybgood]
    #14329900 - 04/21/11 08:38 PM (12 years, 9 months ago)

Quote:

shroomybgood said:
Will a GB be sufficient for doing the transfers?




You can do a lot of things in a good GB with good Techinique:wink:











Glad to see you trying your hand at agar,
if you havent already checked it out heres the link I started out using Grocery Store Agar Tek

I love this Tek and has never failed me,
you can reduce the amount of potato flakes used in the tek to 1/2.



:sunny:


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Invisibleshroomybgood
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Re: Best way to isolate AA+? [Re: Barakanaten]
    #14331740 - 04/22/11 04:53 AM (12 years, 9 months ago)

Thanks Barakanaten!!! That's very helpful!


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InvisibleBarakanaten
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Re: Best way to isolate AA+? [Re: shroomybgood]
    #14332337 - 04/22/11 09:04 AM (12 years, 9 months ago)

Quote:

shroomybgood said:
Thanks Barakanaten!!! That's very helpful!





:datass: I try :blush:


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Invisibleshroomybgood
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Re: Best way to isolate AA+? [Re: Barakanaten]
    #14333414 - 04/22/11 01:32 PM (12 years, 9 months ago)

Alright, I just want to make sure I've got this right? Once I've got a good solid hyphae then I just juice the jar/dish with sterilized water, suck that back up and that works basically like a spore syringe? Would it be best at that point to just noc 1 jar and do GTG for the rest or can I go ahead and noc up several jars and be confident the myc will be solid?


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InvisibleBarakanaten
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Re: Best way to isolate AA+? [Re: shroomybgood]
    #14334466 - 04/22/11 05:40 PM (12 years, 9 months ago)

Quote:

shroomybgood said:
Alright, I just want to make sure I've got this right? Once I've got a good solid hyphae then I just juice the jar/dish with sterilized water, suck that back up and that works basically like a spore syringe? Would it be best at that point to just noc 1 jar and do GTG for the rest or can I go ahead and noc up several jars and be confident the myc will be solid?





you can go either route, thats gonna be up to
you and why you want to do what you want to do.

I like to take an extra dish and cut it into a bunch of chunks,
toss right into a grain jar and shake right away.
Its super fast this way, also your adding tonnes of nutes to the jar.
Generally my jars done this way are 100% in 5 days.

then ffrom there I g2g into 4-10 jars:super:

LC's and Glc's are both great and have their place just like anything else.

also keep track of what generation of spawn your on when you g2g,
its kind of like isolating in a way if you do it from MS.

I personally like the whole G2G thing,
but thats mostly because I am terribe at LC's and GLC's are easy and proven...


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Invisiblemister
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Re: Best way to isolate AA+? [Re: Barakanaten]
    #14334702 - 04/22/11 06:43 PM (12 years, 9 months ago)

If you are going to be doing any agar work and are going to be growing for a long period of time. then invest in a flowhood. the glove box will quickly be a pain in the ass and not give you the freedom of movement you will want or the best chance of a sterile working environment...as you seen from the set of pictures above with the big spot of mold on one of the plates done with a glove box. RR is right though. aim for consistant results to which more fruits mature, rather than abort. That is just wasted substrate that could of been used better.

Congrats on your progress of the AA+.


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OfflineEctius
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Re: Best way to isolate AA+? [Re: Barakanaten]
    #14335285 - 04/22/11 08:44 PM (12 years, 9 months ago)

Quote:

Barakanaten said:
Quote:

shroomybgood said:
Will a GB be sufficient for doing the transfers?




You can do a lot of things in a good GB with good Techinique;)



Holy Organization, Batman!

Nice lemon juice squeezer, btw.  :laugh:


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InvisibleBarakanaten
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Re: Best way to isolate AA+? [Re: mister]
    #14335352 - 04/22/11 08:54 PM (12 years, 9 months ago)

Quote:

mister said:
If you are going to be doing any agar work and are going to be growing for a long period of time. then invest in a flowhood. the glove box will quickly be a pain in the ass and not give you the freedom of movement you will want or the best chance of a sterile working environment...as you seen from the set of pictures above with the big spot of mold on one of the plates done with a glove box. RR is right though. aim for consistant results to which more fruits mature, rather than abort. That is just wasted substrate that could of been used better.

Congrats on your progress of the AA+.





this is all true but do what you can for now.

for the record...
lol the dish above was contaminated from a filter
that got tore off and exposed to unsteril conditions...lol


that nice patch of Black mold was hiding behind my lab assistants litter box.

I am actually about 3 contams out of 60+ dishes in the GB...lol
funny thing though I didnt get the contams from the GB untill I was more experienced,
Perhaps I was more comfortable and threw a bit of caution to the wind.

-Ectius yeah that lemon bottle makes a good wash bottle...:lol:
its fun to improvise and adapt.:grin:

:sunny:


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Invisibleshroomybgood
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Re: Best way to isolate AA+? [Re: Barakanaten]
    #14337413 - 04/23/11 08:12 AM (12 years, 9 months ago)

Quote:

Barakanaten said:
Quote:

shroomybgood said:
Alright, I just want to make sure I've got this right? Once I've got a good solid hyphae then I just juice the jar/dish with sterilized water, suck that back up and that works basically like a spore syringe? Would it be best at that point to just noc 1 jar and do GTG for the rest or can I go ahead and noc up several jars and be confident the myc will be solid?





you can go either route, thats gonna be up to
you and why you want to do what you want to do.

I like to take an extra dish and cut it into a bunch of chunks,
toss right into a grain jar and shake right away.
Its super fast this way, also your adding tonnes of nutes to the jar.
Generally my jars done this way are 100% in 5 days.

then ffrom there I g2g into 4-10 jars:super:

LC's and Glc's are both great and have their place just like anything else.

also keep track of what generation of spawn your on when you g2g,
its kind of like isolating in a way if you do it from MS.

I personally like the whole G2G thing,
but thats mostly because I am terribe at LC's and GLC's are easy and proven...




When you do this technique, do you use the entire contents of one jar/dish to one quart? Thanks for the info and replies guys! I almost feel like I'm in school again only I like the subject! lol


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Invisibleshroomybgood
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Re: Best way to isolate AA+? [Re: shroomybgood]
    #14361906 - 04/27/11 03:59 PM (12 years, 9 months ago)

Nevermind the above question as it was answered in a post above. lol I am soaking up so much information, I think I get slightly overloaded at times. Finally found agar today so I'm about to start mixing it up. Can I just store the unused poured dishes in the fridge? Should I put them in bags?


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InvisibleBarakanaten
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Re: Best way to isolate AA+? [Re: shroomybgood]
    #14362301 - 04/27/11 05:01 PM (12 years, 9 months ago)

Quote:

shroomybgood said:
Can I just store the unused poured dishes in the fridge? Should I put them in bags?




Thats a tough one, the refrigerator can be a huge vector
for contaminates. I personally havent had an issue with
them in the refrigerator, but I know a lot of folks who
have caught hell trying to figure out where their contam
vector is and it ended up being the fridge. The other side
of that coin is many folks get away with it as well. So this
leads me to believe 2 things:

1.keep your fridge clean
2.be sure all your filter material/GE Filters are quality.

I would assume if you use parafilm or something
equivalent it shouldn't be an issue for your plates.



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Invisibleshroomybgood
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Re: Best way to isolate AA+? [Re: Barakanaten]
    #14362971 - 04/27/11 06:58 PM (12 years, 9 months ago)

Alright so I did the grocery store agar tek that you linked to me ^^^. I used the same jars and everything exactly the same way. They are cooling at the moment and my plan was to put them in ziplock bags and stick them in the fridge. Do I need to run out and but parafilm too? It's not a problem if that's what I need to do.


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Offlinesteelmonkey
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Re: Best way to isolate AA+? [Re: shroomybgood]
    #14363007 - 04/27/11 07:04 PM (12 years, 9 months ago)

Ive never used it...either a zip-loc sandwich bag or saran wrap has always worked for me but thats just my opinion lots of people use parrafin wax


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InvisibleBarakanaten
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Re: Best way to isolate AA+? [Re: steelmonkey]
    #14363570 - 04/27/11 09:03 PM (12 years, 9 months ago)

Quote:

shroomybgood said:
Alright so I did the grocery store agar tek that you linked to me ^^^. I used the same jars and everything exactly the same way. They are cooling at the moment and my plan was to put them in ziplock bags and stick them in the fridge. Do I need to run out and but parafilm too? It's not a problem if that's what I need to do.




Really its a personal preference I guess,
I could scare you and say "ah man thats not going to work"
Then you might try it anyways and it might work and you may
say to yourself "that guy's a ding dong"...:lol:

I'll give you an example:

I used 50% coffee for my monos Moarrrrr Coffee Mono and used Damions coir tek

well the responses I got when I announced I used 50% SCG Spent Coffee Grinds
and a cup or two of straight coffee was :whoa: "good luck with that"  ":sarcasticclap:" ...lol
Also some said that the coir tek might only be good for a flush or two...before it contams.

in the end they all fruited fairly fast and beautiful
despite a few n00b mistakes and I brought the tubs to
5 flushes and tossed them out and called it a day.

If it were me I would go ahead and use the ziplocks or gladwrap
and invest in some parafilm as soon as you can, its just one more
step towards solidifying you as an amateur mycologist and one more
thing that you can eliminate as suspect when something goes awry.

Also remember that the temperature fluctuations in a regular "home" refrigerator
can damage cultures during long term storage, I have heard RR uses an insulated
lunch cooler/box thingy to keep the temps stable and preserve the culture for long term storage.


:sunny:


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Edited by Barakanaten (04/27/11 09:17 PM)


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Invisibleshroomybgood
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Re: Best way to isolate AA+? [Re: Barakanaten]
    #14363624 - 04/27/11 09:12 PM (12 years, 9 months ago)

Cool. I will be doing my first transfer in the morning. I'm pretty stoked about this part of my mycology experience. I'm really looking forward to seeing the results of isolation.


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Re: Best way to isolate AA+? [Re: shroomybgood]
    #14363983 - 04/27/11 10:12 PM (12 years, 9 months ago)

Well good luck I hope it goes well for you its always nice to see someone trying agar


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Re: Best way to isolate AA+? [Re: steelmonkey]
    #14406756 - 05/05/11 06:43 PM (12 years, 8 months ago)

So for my first I used MS from some syringes. They are taking off now so I plan on doing another transfer tomorrow. I also transfered some tissue from a second flush victim. I wasn't sure about using flesh from a second but I really had no other choice other than to not do it at all. Can I still get good results that way?


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Re: Best way to isolate AA+? [Re: shroomybgood]
    #14406778 - 05/05/11 06:47 PM (12 years, 8 months ago)

Its possible,how did your second flush go?


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Re: Best way to isolate AA+? [Re: steelmonkey]
    #14406787 - 05/05/11 06:48 PM (12 years, 8 months ago)

First was good and second even better!


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Re: Best way to isolate AA+? [Re: shroomybgood]
    #14407452 - 05/05/11 09:04 PM (12 years, 8 months ago)

You might experience slower growth than from a first flush pin but if you do say a three transfer isolate I bet it all turns out great


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Re: Best way to isolate AA+? [Re: steelmonkey]
    #14423465 - 05/09/11 07:38 AM (12 years, 8 months ago)

So far I've got myc on about 10 different agar jars. Only 1 has noticeable contamination. I'm debating just tossing that jar? I know I can isolate away from the contamination but I'm just a little nervous about it.


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Re: Best way to isolate AA+? [Re: shroomybgood]
    #14424374 - 05/09/11 12:21 PM (12 years, 8 months ago)

isolating from contamination is an advanced technique. I would toss it for now and just keep on practicing. After a while you will feel good about it and want to try something like that.


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Re: Best way to isolate AA+? [Re: sandman420]
    #14424444 - 05/09/11 12:38 PM (12 years, 8 months ago)

Or just keep it seperated from the rest and try your hand at it and see how you do,at the very least you will learn what not to do


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Invisibleshroomybgood
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Re: Best way to isolate AA+? [Re: steelmonkey]
    #14424466 - 05/09/11 12:42 PM (12 years, 8 months ago)

After checking this afternoon I found that 1 more has contaminated. I will try my hand at getting some good myc out of them. I'm going to have to learn sometime?


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Invisibleshroomybgood
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Re: Best way to isolate AA+? [Re: shroomybgood]
    #14436210 - 05/11/11 06:12 PM (12 years, 8 months ago)

Well I did the transfers 1 at a time and kept them away from everything else. So far they are doing fine and I'm watching closely for signs of contam. Agar work does seem to be pretty tedious but I'm hoping the results make it worth it.


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Invisibleshroomybgood
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Re: Best way to isolate AA+? [Re: shroomybgood]
    #14470687 - 05/17/11 10:53 PM (12 years, 8 months ago)

Little update. I'm kind of proud of my sterile practice and "good luck". The transfers from the contaminated agar jars are a success! I will be transferring one more time tomorrow and hopefully I'll have what I'm looking for. I'm super anxious to see the resulting fruits. Thanks for all the advice!


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"GET IT TOGETHER ONE MORE TIME"


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Offlinesteelmonkey
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Re: Best way to isolate AA+? [Re: shroomybgood]
    #14471763 - 05/18/11 04:47 AM (12 years, 8 months ago)

Its always so exciting to see your isolate fruit and see if you did everything right,the wait is a killer.Got any pics?


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Invisibleshroomybgood
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Re: Best way to isolate AA+? [Re: steelmonkey]
    #14471960 - 05/18/11 07:07 AM (12 years, 8 months ago)

I have a long answer to that short question. I once bought a camera to take pictures of the cars I worked on. Put it in my tool box and as I would be watching the cars leave I'd say to myself, "Damn I should've got a picture". I am a believer in forward motion and leaving the past behind. Memories are good enough for me even though mine sucks. I understand the point in taking pics of our progress and sharing our experience with others but I'm not going to buy a camera for this alone. That being said, also I lurked on here for a year almost before I got the balls to get an account. Pictures are still far off! lol


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Offlinesteelmonkey
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Re: Best way to isolate AA+? [Re: shroomybgood]
    #14472014 - 05/18/11 07:36 AM (12 years, 8 months ago)

I don't take many pics myself just because I'm not a photography type of guy but recently I came across a few pics of my first ever fruiting cake from god maybe 6 years ago and it warmed my heart


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Offlineilladelph
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Re: Best way to isolate AA+? [Re: m_jacobs]
    #27623759 - 01/18/22 04:18 PM (2 years, 11 days ago)

i know this post is a decade old; however, I would like to ask a question/clear something up. step 4 in your process:
"Then go Ager to small jar of WBS.  Once WBS is 100%, I would do GLC."
may I ask why you add this step instead of simply going straight from agar to LC. by now you know that your cultures clean right...? hopefully theres something I'm missing and someone can teach me something. or shit has just changed, it has been 10 years lol. thnx guys


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β€œall fungi are edible,
some fungi are only edible once.”
― Terry Pratchett
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"two possibilities exist:
either we are alone in the universe or we are not...
both are equally terrifying".
― Arthur C. Clark
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---together we stand, divided we fall---


Edited by illadelph (01/18/22 04:38 PM)


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OfflineMetaSophia
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Re: Best way to isolate AA+? [Re: illadelph]
    #27623949 - 01/18/22 08:04 PM (2 years, 11 days ago)



I think you're going to need to learn to live in the light of the mystery, friend.


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