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OfflineKillerclowns87
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agar questions
    #24279613 - 04/28/17 07:12 PM (7 years, 8 months ago)

Good evening everyone, hope all is well!

So ive been messing with agar for the last few months, and of course just like everyone else i love it. However...


For one... labeling...

I keep a notebook and record everything that i do and observe in my spawn, bulk, and agar but i have trouble matching my notes to its correspondence sometimes... need a good labeling system.

next... all of these plates!!

How i have been conducting things is.. i swab a print.. put that to agar and make transfers until i get what i think is "a good looking culture" i'm hoping i am getting somewhere close to a "mono culture".

Then i will cut that plate into quarters and noc quart jars with 3/4 of the plate and refrigerate the last 1/4 (these are properly labeled, incase i want to come back to a culture that did really well in bulk) heres what i got so far for transfers/plates.



These are just a couple.. i'm having trouble selecting which plate and which sectors, there are soo many!! I got serious and ordered a case of plates and plenty of agar,
i have the resources.. just... overwhelmed!

I'd like to start with this question if i may.. in this plate:

What exactly does this type of mycelium entail? The strands are darker and more define and and thicker in some spots. Like that one "poker" @ 5oclock

And those 4 well defined guys @ 9-9:30. you see them?



Compared to a "normal" looking plate such as:

Could someone share with me their normal "agar routine"?
Do you use agar kind of like i do?

spore > agar > transfers to get even, parallel, no sectors growth > grain > bulk??

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Invisiblejohn_conner

Registered: 02/21/17
Posts: 243
Re: agar questions [Re: Killerclowns87]
    #24279662 - 04/28/17 07:35 PM (7 years, 8 months ago)

though I don't have any input at the moment I will however be watching this closely:takingnotes: as I am in the same boat as you just not as far along. Keep up the good work! looking good

Edited by john_conner (04/28/17 07:36 PM)

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InvisibleGerms
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Re: agar questions [Re: john_conner]
    #24279719 - 04/28/17 07:55 PM (7 years, 8 months ago)

What recipe was the 5th pic made from?


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InvisibleTheMadHatter420
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Re: agar questions [Re: Germs]
    #24279735 - 04/28/17 08:00 PM (7 years, 8 months ago)

If that is from spore, then you are far far from mono culture. Mono cultures usually result from a clone, that's made up of possibly a couple dozen mono cultures itself, being cleaned up and isolated. There isn't really any reason for a mono culture if all you want is fruits.

Clean it up, grow it out, pick a GOOD clone, clean that up and have good, fairly consistent genetics. If you REALLY want a mono culture, you go from there, but each mono culture has to be tested. It could end up being a dud.


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Invisiblec10h12n2o
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Re: agar questions [Re: Germs]
    #24279739 - 04/28/17 08:02 PM (7 years, 8 months ago)

I'm in the middle of a ton of agar work as we speak, I will post shortly with details of some techniques that I think will help you out :smile:

Also will answer your questions and give feedback on your plates


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

C10's Agar Guide + Tips and Tricks | c10's Flow Hood Build Guide


"Partial knowledge is more triumphant than complete knowledge; it takes things to be simpler than they are, and so makes its theory more popular and convincing."

"Convictions are more dangerous enemies of truth than lies"
― Friedrich Nietzsche

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OfflineOrganic_Magic
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Re: agar questions [Re: Germs]
    #24279763 - 04/28/17 08:11 PM (7 years, 8 months ago)

I do it like this except I hate putting spores to agar as I have trouble with germination rates but whatever. I just take tissue clones or grain spawn to agar then transfer etc.

Those plates all look really nice and are showing some very defined sectors. I would not try for a mono-culture per say, but lean more toward finding a consistent culture that's a fast colonizer (and good fruiter). Finding a good mono-culture can take literally thousands of transfers and then it might not even perform well.

Just try and take as small of transfers as possible from the outermost leading edge to "capture" that fast/aggressive mycelium.
:super:


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OfflineKillerclowns87
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Re: agar questions [Re: c10h12n2o]
    #24279784 - 04/28/17 08:17 PM (7 years, 8 months ago)

Quote:

c10h12n2o said:
I'm in the middle of a ton of agar work as we speak, I will post shortly with details of some techniques that I think will help you out :smile:

Also will answer your questions and give feedback on your plates




you are literally the man!! You keep coming with goodies!!

I was going to post more in there speaking of how i am afraid to crack open a spore print "i recently got from a giveaway":thumbup::heart:. But i thought my post was getting lengthy already.

Thanks again bro!

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OfflineKillerclowns87
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Re: agar questions [Re: Organic_Magic]
    #24279799 - 04/28/17 08:23 PM (7 years, 8 months ago)

Quote:

Organic_Magic said:

Those plates all look really nice and are showing some very defined sectors. I would not try for a mono-culture per say, but lean more toward finding a consistent culture that's a fast colonizer (and good fruiter). Finding a good mono-culture can take literally thousands of transfers and then it might not even perform well.

Just try and take as small of transfers as possible from the outermost leading edge to "capture" that fast/aggressive mycelium.
:super:





Good info, thank you for that.

As simple as it was you've already eased my stress and a lot of confusion.:thumbup:

Basically that's what i have been doing thinking i had a monoculture when there were absolutely no more sectors and everything was nice and even. Although i now know different, at least i was still heading in the right direction. Good stuff!

Thanks again:grin:

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InvisibleBugsRucker
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Re: agar questions [Re: TheMadHatter420]
    #24279802 - 04/28/17 08:24 PM (7 years, 8 months ago)

agar is a great resource.

to grow mycelium.

only mycelium. clean with no molds or bacteria.

in my most humble of opinion there is way too much talk about isolating and isolates and monocultures and separating the hippocampus from the pineal gland etc.

do not doubt for one second that agar is great for growing ultra clean mycelium, and its stupid easy. EDIT: AND YOU OBVIOUSLY HAVE THE HANG OF IT, NICE LOOKING PLATES.

that is more than most of us amateur cultivators will ever need.

creating a single isolate or mono culture is 100x more work and transfers than it takes to grow nice and clean mycelium on a plate.

fruiting and testing a bunch of isolates to find one that is significantly superior to just using clean mycelium is 100x more work and transfers than creating the single isolate mentioned above.

mostly i think its just semantics and terms being introduced without being completely understood and thats not a huge deal but agar for clean mycelium is a killer tool.

cultivating and identifying killer isolates tho is more like its own hobby in itself, similar to if you were trying to create new varieties by cross breeding with rattlesnake venom or something.

rant over and nice agar work op. no disrespect intended at all.

as far as labeling goes... ugh, i still am not sure. but i start with syringes so i label my first one 'ms' (multispore), then my transfer from that i label '1' and my transfer from 1 i label '2' and so on. when i am happy with the growth and want to get a bunch of plates i would take a bunch of transfers from a single plate and label them all '2.1' so i know they are all transfers from the second 'generation' from my original multispore plate. hope it helps.

Edited by BugsRucker (04/28/17 08:26 PM)

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OfflineKillerclowns87
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Re: agar questions [Re: Germs]
    #24279844 - 04/28/17 08:43 PM (7 years, 8 months ago)

Quote:

Germs said:
What recipe was the 5th pic made from?





They are all made from same recipe : a basic pda recipe
                                                        7g Dextrose
                                                        9g agar
                                                        5g potato flakes
                                                        : 500ml

Edited by Killerclowns87 (04/28/17 09:24 PM)

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Invisiblec10h12n2o
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Re: agar questions [Re: Killerclowns87] * 1
    #24280269 - 04/28/17 11:54 PM (7 years, 8 months ago)

glad to help my friend :smile:

i gotta say, you are WAY on the right track! you are doing lots of things the right way, and seem to have done your research, are asking the right questions, and thinking critically,  which pretty much means you are gonna kick ass at cultivation :smile:

just the fact that we dont have to spend 10 threads trying to talk you out of some bullshit you learned from a kit is a huge advantage :lol:

as far as labeling, i write a number for each race/variety, and "c" for clones (often plus a designation), write the date, and "x-10" for the 10th transfer, something like "33c-Mon.  x-12  1/10" would mean AA+ clone variant "monster" (weighed 295 grams) and on the 12th transfer, made on january 10th

here is a random one:


when i get to a culture i want to use (usually a highly organized culture) i will assign a string of numbers and letters to it (33A2b for instance) and keep a sample of it, and use it to designate all projects using that culture

i track all this in spreadsheets where i keep my notes. my handwriting is shit. i also write directly on the plates with sharpies, usually i use red marker for clones and black for MS, but need a new red marker lol...

as far as your plates:

they look great, you are doing a great job :smile: but as others have said, you really dont need isolates for your purposes, and they can often take an insane amount of work to get.... sometimes a culture will look perfectly round for 10 transfers and then show a dozen sectors, its crazy... its really tricky getting strains and strain groups to distinguish themselves sometimes

you should really just focus on getting clean, aggressive, organized cultures at this point, thats the ticket :smile:

just keep transferring the leading edge of the most aggressive, healthiest looking myc until you get something really organized

so here is a little trick that i DIG: its basically a serial dilution right on the surface of the agar



by streaking spores in this way, you have LOTS of colonies to choose from, with far fewer strains per colony than a typical blob of spores would result in. this can allow you to get an isolate in way less transfers especially if you use the later zones (4, 5), and is also very useful in cleaning up prints, and gives hundreds of possible colonies to choose from

my technique:

i use MEA almost exclusively, only occasionally using PDA or MYPA to switch things up

my recipe: 20g agar, 15g light malt extract, 1L water

i fill 2 1 L media bottles halfway full (to avoid boiling over) with the lids barely cracked  and load them in the PC at an angle so they will fit. at the same time i sterilize 4 scalpel handles in individual foil envelopes

when i open the PC, i immediately tighten the lids. when the bottles read 60 degrees C on an infrared thermometer, i begin to pour them (before they hit 50), usually about 60-80 90mm plates per L of agar. i often have to swirl the stack of plates a bit to coat the bottom of the plate, trying to use as little as possible (probably bad technique, but i prefer it thin, so that i dont have to trim my transfers

after i finish pouring, i either bag them up in gallon freezer bags or use them immediately

i keep 100pc scalpel blades handy, and use a fresh one for each session. i also use a combination of an alcohol lamp and a butane bunsen burner ( i LOVE this thing, perfect for hands free dab prep, or myco stuff :smile: ), i use the alcohol to light the butane burner, just so i dont wear out the piezo ignition. if i wasnt doing such huge volumes i would just leave the butane bunsen running



i used to use face masks, but my face always got hot and sweaty, now i use a face shield and love it



when using a SAB it can make a BIG difference to use a damp towel in the bottom (changed each session) and work on a cookie rack a few inches above the towel. this traps contams and moves your workspace above the dirty surface

it can also be handy to have a light under that rack, for back lighting plates and picking out features you want to transfer or avoid

i also LOVE parafilm, which i cut by drawing out about 5 feet, cutting off 2.5 feet, and clamping it so that it is 2 layers thick, then using a retractable scalpel to cut 3/4 inch strips (2 at a time) and form a large stack of these parafilm wrappers before the session, so they are handy and ready to go when i need them

i also use put each plate inside a ziploc sandwich baggie, just for an extra layer of protection, so that i can handle it out in the open while it is culturing

i keep a shaving towel in a freezer ziploc, and take it out to wipe my gloves and forearms as needed. the bag keeps the alcohol from evaporating as fast so it lasts much longer

let me know if you have any questions about any of that :smile: looking forward to seeing your future projects :smile:


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

C10's Agar Guide + Tips and Tricks | c10's Flow Hood Build Guide


"Partial knowledge is more triumphant than complete knowledge; it takes things to be simpler than they are, and so makes its theory more popular and convincing."

"Convictions are more dangerous enemies of truth than lies"
― Friedrich Nietzsche

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InvisibleTripping2Adventure
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Re: agar questions [Re: c10h12n2o]
    #24280483 - 04/29/17 02:26 AM (7 years, 8 months ago)

Quote:

c10h12n2o said:
glad to help my friend :smile:

i gotta say, you are WAY on the right track! you are doing lots of things the right way, and seem to have done your research, are asking the right questions, and thinking critically,  which pretty much means you are gonna kick ass at cultivation :smile:

just the fact that we dont have to spend 10 threads trying to talk you out of some bullshit you learned from a kit is a huge advantage :lol:

as far as labeling, i write a number for each race/variety, and "c" for clones (often plus a designation), write the date, and "x-10" for the 10th transfer, something like "33c-Mon.  x-12  1/10" would mean AA+ clone variant "monster" (weighed 295 grams) and on the 12th transfer, made on january 10th

here is a random one:


when i get to a culture i want to use (usually a highly organized culture) i will assign a string of numbers and letters to it (33A2b for instance) and keep a sample of it, and use it to designate all projects using that culture

i track all this in spreadsheets where i keep my notes. my handwriting is shit. i also write directly on the plates with sharpies, usually i use red marker for clones and black for MS, but need a new red marker lol...

as far as your plates:

they look great, you are doing a great job :smile: but as others have said, you really dont need isolates for your purposes, and they can often take an insane amount of work to get.... sometimes a culture will look perfectly round for 10 transfers and then show a dozen sectors, its crazy... its really tricky getting strains and strain groups to distinguish themselves sometimes

you should really just focus on getting clean, aggressive, organized cultures at this point, thats the ticket :smile:

just keep transferring the leading edge of the most aggressive, healthiest looking myc until you get something really organized

so here is a little trick that i DIG: its basically a serial dilution right on the surface of the agar



by streaking spores in this way, you have LOTS of colonies to choose from, with far fewer strains per colony than a typical blob of spores would result in. this can allow you to get an isolate in way less transfers especially if you use the later zones (4, 5), and is also very useful in cleaning up prints, and gives hundreds of possible colonies to choose from

my technique:

i use MEA almost exclusively, only occasionally using PDA or MYPA to switch things up

my recipe: 20g agar, 15g light malt extract, 1L water

i fill 2 1 L media bottles halfway full (to avoid boiling over) with the lids barely cracked  and load them in the PC at an angle so they will fit. at the same time i sterilize 4 scalpel handles in individual foil envelopes

when i open the PC, i immediately tighten the lids. when the bottles read 60 degrees C on an infrared thermometer, i begin to pour them (before they hit 50), usually about 60-80 90mm plates per L of agar. i often have to swirl the stack of plates a bit to coat the bottom of the plate, trying to use as little as possible (probably bad technique, but i prefer it thin, so that i dont have to trim my transfers

after i finish pouring, i either bag them up in gallon freezer bags or use them immediately

i keep 100pc scalpel blades handy, and use a fresh one for each session. i also use a combination of an alcohol lamp and a butane bunsen burner ( i LOVE this thing, perfect for hands free dab prep, or myco stuff :smile: ), i use the alcohol to light the butane burner, just so i dont wear out the piezo ignition. if i wasnt doing such huge volumes i would just leave the butane bunsen running



i used to use face masks, but my face always got hot and sweaty, now i use a face shield and love it



when using a SAB it can make a BIG difference to use a damp towel in the bottom (changed each session) and work on a cookie rack a few inches above the towel. this traps contams and moves your workspace above the dirty surface

it can also be handy to have a light under that rack, for back lighting plates and picking out features you want to transfer or avoid

i also LOVE parafilm, which i cut by drawing out about 5 feet, cutting off 2.5 feet, and clamping it so that it is 2 layers thick, then using a retractable scalpel to cut 3/4 inch strips (2 at a time) and form a large stack of these parafilm wrappers before the session, so they are handy and ready to go when i need them

i also use put each plate inside a ziploc sandwich baggie, just for an extra layer of protection, so that i can handle it out in the open while it is culturing

i keep a shaving towel in a freezer ziploc, and take it out to wipe my gloves and forearms as needed. the bag keeps the alcohol from evaporating as fast so it lasts much longer

let me know if you have any questions about any of that :smile: looking forward to seeing your future projects :smile:




So many useful techniques. Glad I hopped on this thread, posting to keep this info:thumbup:

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Offlinedhype773
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Re: agar questions [Re: c10h12n2o]
    #24280627 - 04/29/17 05:40 AM (7 years, 8 months ago)

Quote:

c10h12n2o said:
glad to help my friend :smile:

i gotta say, you are WAY on the right track! you are doing lots of things the right way, and seem to have done your research, are asking the right questions, and thinking critically,  which pretty much means you are gonna kick ass at cultivation :smile:

just the fact that we dont have to spend 10 threads trying to talk you out of some bullshit you learned from a kit is a huge advantage :lol:

as far as labeling, i write a number for each race/variety, and "c" for clones (often plus a designation), write the date, and "x-10" for the 10th transfer, something like "33c-Mon.  x-12  1/10" would mean AA+ clone variant "monster" (weighed 295 grams) and on the 12th transfer, made on january 10th

here is a random one:


when i get to a culture i want to use (usually a highly organized culture) i will assign a string of numbers and letters to it (33A2b for instance) and keep a sample of it, and use it to designate all projects using that culture

i track all this in spreadsheets where i keep my notes. my handwriting is shit. i also write directly on the plates with sharpies, usually i use red marker for clones and black for MS, but need a new red marker lol...

as far as your plates:

they look great, you are doing a great job :smile: but as others have said, you really dont need isolates for your purposes, and they can often take an insane amount of work to get.... sometimes a culture will look perfectly round for 10 transfers and then show a dozen sectors, its crazy... its really tricky getting strains and strain groups to distinguish themselves sometimes

you should really just focus on getting clean, aggressive, organized cultures at this point, thats the ticket :smile:

just keep transferring the leading edge of the most aggressive, healthiest looking myc until you get something really organized

so here is a little trick that i DIG: its basically a serial dilution right on the surface of the agar



by streaking spores in this way, you have LOTS of colonies to choose from, with far fewer strains per colony than a typical blob of spores would result in. this can allow you to get an isolate in way less transfers especially if you use the later zones (4, 5), and is also very useful in cleaning up prints, and gives hundreds of possible colonies to choose from

my technique:

i use MEA almost exclusively, only occasionally using PDA or MYPA to switch things up

my recipe: 20g agar, 15g light malt extract, 1L water

i fill 2 1 L media bottles halfway full (to avoid boiling over) with the lids barely cracked  and load them in the PC at an angle so they will fit. at the same time i sterilize 4 scalpel handles in individual foil envelopes

when i open the PC, i immediately tighten the lids. when the bottles read 60 degrees C on an infrared thermometer, i begin to pour them (before they hit 50), usually about 60-80 90mm plates per L of agar. i often have to swirl the stack of plates a bit to coat the bottom of the plate, trying to use as little as possible (probably bad technique, but i prefer it thin, so that i dont have to trim my transfers

after i finish pouring, i either bag them up in gallon freezer bags or use them immediately

i keep 100pc scalpel blades handy, and use a fresh one for each session. i also use a combination of an alcohol lamp and a butane bunsen burner ( i LOVE this thing, perfect for hands free dab prep, or myco stuff :smile: ), i use the alcohol to light the butane burner, just so i dont wear out the piezo ignition. if i wasnt doing such huge volumes i would just leave the butane bunsen running



i used to use face masks, but my face always got hot and sweaty, now i use a face shield and love it



when using a SAB it can make a BIG difference to use a damp towel in the bottom (changed each session) and work on a cookie rack a few inches above the towel. this traps contams and moves your workspace above the dirty surface

it can also be handy to have a light under that rack, for back lighting plates and picking out features you want to transfer or avoid

i also LOVE parafilm, which i cut by drawing out about 5 feet, cutting off 2.5 feet, and clamping it so that it is 2 layers thick, then using a retractable scalpel to cut 3/4 inch strips (2 at a time) and form a large stack of these parafilm wrappers before the session, so they are handy and ready to go when i need them

i also use put each plate inside a ziploc sandwich baggie, just for an extra layer of protection, so that i can handle it out in the open while it is culturing

i keep a shaving towel in a freezer ziploc, and take it out to wipe my gloves and forearms as needed. the bag keeps the alcohol from evaporating as fast so it lasts much longer

let me know if you have any questions about any of that :smile: looking forward to seeing your future projects :smile:




:breakthrough:

Wow!  My SAB technique changed the instant I read this!


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

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OfflineCapnZ
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Re: agar questions [Re: dhype773]
    #24280648 - 04/29/17 06:04 AM (7 years, 8 months ago)

Sounds like a lot of us are going thru the same process in about the same place. Here are 2 varieties that are on transfer plates. Decent, but I just ransferred these again picking what I thought was the best  area and sampling out a rice grain sized slice. Just did that Thu night. I use a pretty simple labeling system but it works well for me, even with my plate count growing exponentially. I can't stop!!!
Hello, agar addiction help line??......



--------------------
Deep into the darkness peering, long I stood there wondering, fearing, doubting, dreaming dreams no mortal ever dared to dream before...

Edited by CapnZ (04/29/17 07:44 AM)

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Invisiblec10h12n2o
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Re: agar questions [Re: CapnZ]
    #24280660 - 04/29/17 06:28 AM (7 years, 8 months ago)

I'm glad y'all found it useful :smile:

I been planning to do an agar tips thread for a while now, just been busy


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

C10's Agar Guide + Tips and Tricks | c10's Flow Hood Build Guide


"Partial knowledge is more triumphant than complete knowledge; it takes things to be simpler than they are, and so makes its theory more popular and convincing."

"Convictions are more dangerous enemies of truth than lies"
― Friedrich Nietzsche

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Offlinedhype773
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Re: agar questions [Re: c10h12n2o]
    #24280662 - 04/29/17 06:31 AM (7 years, 8 months ago)

Quote:

c10h12n2o said:
I'm glad y'all found it useful :smile:

I been planning to do an agar tips thread for a while now, just been busy



You should definitely get on that! That's the second bit of agar above from you that has blown my mind!


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OfflineCapnZ
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Re: agar questions [Re: dhype773]
    #24280720 - 04/29/17 07:30 AM (7 years, 8 months ago)

I've been having really good luck with grain water and this stuff



My local Asian store does not carry telephone but they have this brand. For 500 ml:

200 ml grain water ( from oat prep)
300 ml distilled water
2 grams LME
1 pack of the above

That pours 20 plates.

I use a propane torch on low to flame - quick to heat to red hot. Yup change scalpel blades foe every session.

C10 - frigging love the idea of a face shield. Thanks for the tip, I'm getting one!


--------------------
Deep into the darkness peering, long I stood there wondering, fearing, doubting, dreaming dreams no mortal ever dared to dream before...

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OfflineKillerclowns87
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Re: agar questions [Re: c10h12n2o]
    #24280828 - 04/29/17 08:53 AM (7 years, 8 months ago)

Terrific!

I want to thank everyone for all the information here.

I definitely know which way to turn now thanks to everyone here (especially you c102h12n2o!!). I will keep this post updated as i have decided to put all the plates into long term storage(refrigerator) except for one plate:

I plan on sectoring this guy and trying each quart individually in a small 6qt tub, until i get more comfortable with what my main goal is if i'm shooting quality. (weight, potency)

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Re: agar questions [Re: Killerclowns87]
    #24280832 - 04/29/17 08:57 AM (7 years, 8 months ago)

That's a purrrdy plate you got there :thumbup:


--------------------
If I ever give out misinformation please inform me so I can have the correct information. :cheers:

Tmethyl said:
Chuck Norris once roundhouse kicked a monotub that wasn't pinning fast enough. The force of the kick rearranged the genetics of the mushrooms, we now call them Penis Envy.

Caps McGee said:
:thumbsup:
Fun part is figuring out what works best for you

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