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Agar Work


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#41 Microbe

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Posted 30 November 2014 - 04:39 PM

Same way you post pictures. I use the Tapatalk app

Really?

#42 CatsAndBats

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Posted 30 November 2014 - 04:42 PM

microbe, kids these days and their intrawebs and hashtagapps or whatever...  :tinfoil:


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#43 CatsAndBats

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Posted 30 November 2014 - 04:43 PM

hashbrownSelfie!

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Edited by catattack, 30 November 2014 - 04:44 PM.

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#44 Microbe

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Posted 01 December 2014 - 08:56 PM

2 plates that received the TEX plate 1-1 transfer after 24 hours already taking over the plate. I expect these to be colonized in 4 days but we will see.

498d54cafe778c1404cf2f3858e81eaf.jpg

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This is one of the plates that was smeared smeared with the culture from the master slant. This is the proof its not a mono but dont be fooled by this as its a great specimen and will be very rhizomorphic after growing out through a single quart of spawn. I would put this into grain any day. I will not isolate this as i already working on isolating a culture from the same slant.

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The point im trying to make here by the crappy picture as i gave up after several attempts to capture a clear one, is that after 24 hours of the transfer it is recovered starting to start growing out. You can see though that it is much slower then the first 2 plates of the 1-1 plate i had been working with, that is the TEX strain :)

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My EC plates are really bad and again no where close to being ready for grain! In fact these are garbage IMO. Weird looking also!

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Not very informative but just wanted to share with you.

Edited by Microbe77, 01 December 2014 - 08:59 PM.

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#45 wharfrat

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Posted 01 December 2014 - 09:53 PM

looking good as always brother. thanks for sharing


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#46 gremlinchode

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Posted 01 December 2014 - 11:51 PM

This is awesome thank you so much for sharing.
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#47 invisibilitysyndrome

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Posted 02 December 2014 - 08:15 AM

Hey. I tried pics but had condensation. Ill try today again. My plates are very fluffy no rizo will shoot pics asap

#48 Microbe

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Posted 02 December 2014 - 08:58 AM

Hey. I tried pics but had condensation. Ill try today again. My plates are very fluffy no rizo will shoot pics asap

If you multispore will almost always start off like that. It doesnt mean its a bad fruiter or even slow its just my preference to use mostly myc that is rhizomorphic. I would suggest transferring before the plate grows out, maybe a couple just in case you loose one. Then inoculate some grain and fruit the remaining culture. Then by then you should have a solid culture by then on agar then fruit that beast and more then likely it will preform better but is very possible it under performs but I doubt that it will. Cant wait to see the pics!

Edited by Microbe77, 02 December 2014 - 08:58 AM.


#49 CatsAndBats

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Posted 02 December 2014 - 11:11 AM

Mic, side note.  So far so good on my isolate of the fatty, It's showing white growth (i hope)


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#50 invisibilitysyndrome

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Posted 02 December 2014 - 01:02 PM

20141202_125414.jpg 20141202_125347.jpg 20141202_125301.jpg 20141202_125239.jpg
All done in sab
W loop and smeared witch might have been a bad idea.
Bplus doing better w center inoc no smear.allothers smeared.
Incubate at 80 degree...pe6 only one no growth but has contam so idk....20141202_125635.jpg

Edited by invisibilitysyndrome, 02 December 2014 - 01:03 PM.


#51 Microbe

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Posted 02 December 2014 - 02:40 PM

attachicon.gif20141202_125635.jpgattachicon.gif20141202_125635.jpgattachicon.gif20141202_125414.jpgattachicon.gif20141202_125347.jpgattachicon.gif20141202_125301.jpgattachicon.gif20141202_125239.jpg
All done in sab
W loop and smeared witch might have been a bad idea.
Bplus doing better w center inoc no smear.allothers smeared.
Incubate at 80 degree...pe6 only one no growth but has contam so idk....attachicon.gif20141202_125635.jpg

Alright you are about to be overwhelmed keeping up with all these plates. I would focus on 1 plate right now so you can give that culture your undivided attention. The B+ plate on the right in your pic that are laid out in a + pattern, has a mysterious spot on it. The first advice if that is what your seeking, is to make sure you keep your inoculate centered regardless of method. It can be challanging at times but it is very important as it can be one of your first signs of a contaminated plate. If you keep it centered and get growth anywhere else it more then likely is a contam. Alot of molds, at least the ones i have seen in my plates looks identical to tomentose mycelium of the Pscilocybe Cubensis myc. Some of these plates have mold on them. I would recommend that you take a plate you want to work with and either fruit the others pending the results of some of the growth that looks suspect to me. Or toss in cold storage. It will be a logistical nightmare to work with all those plate. You would be surprised what the myc can do when allowed to grow out over grain surface. You could always go back to agar with a colonized piece of grain also. If you a direct question about a plate i will give you my advice. Remember i am no expert i just have a sound process and getting very lucky at the selection at times. ToRn2bits put me on a pedestal lol and i am not doing anything that hasnt already been done or that you cant do. Man oh man i wished i would habe documented my first agar work with mushroom mycelium. I cultivated 20 plates of mold in my first session. I seen white growth and was like hell yeah then it turned green. The first step is deciding you are going to do agar work and every single step is critical from preperation of nutrient agar to pouring, to technique when open and closing. But man alive those half dozen of plates are about to be a dozen or more at least assuming you only select from 1 sector. I will post another response momentarily.

#52 Microbe

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Posted 02 December 2014 - 02:44 PM

Mic, side note. So far so good on my isolate of the fatty, It's showing white growth (i hope)

Any pics. I still cant figure out taptalk. As soon as i was getting efficient it was updated and completly changed. I dont get all the alerts and at times someone asks a question and i answer it but then realized that question was asked a month ago. My alert settings are set accordingly on myctopia and with the app. I need to use the mobile version of topia unless im posting pics. But do you have a picture of the clone? Post it here.

#53 Microbe

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Posted 02 December 2014 - 02:46 PM

.

Edited by Microbe77, 02 December 2014 - 02:48 PM.


#54 CatsAndBats

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Posted 02 December 2014 - 02:50 PM

 

Mic, side note. So far so good on my isolate of the fatty, It's showing white growth (i hope)

Any pics. I still cant figure out taptalk. As soon as i was getting efficient it was updated and completly changed. I dont get all the alerts and at times someone asks a question and i answer it but then realized that question was asked a month ago. My alert settings are set accordingly on myctopia and with the app. I need to use the mobile version of topia unless im posting pics. But do you have a picture of the clone? Post it here.

 

Way too small to photograph (it's in a half pint), it's a little bruised piece or two, but there is non-bruised white growth. By next week I will be in a spot to move to agar and dishes..

 

I can't post on tapatalk at all..



#55 Microbe

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Posted 02 December 2014 - 02:53 PM

I promise that i will make a conscious effort to break my ling winded posts up into paragraphs so that it makes it a little more simpler to read them. Now i said effort and at times i may forget ;)

Edited by Microbe77, 02 December 2014 - 02:53 PM.


#56 Microbe

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Posted 02 December 2014 - 03:37 PM

attachicon.gif20141202_125635.jpgattachicon.gif20141202_125635.jpgattachicon.gif20141202_125414.jpgattachicon.gif20141202_125347.jpgattachicon.gif20141202_125301.jpgattachicon.gif20141202_125239.jpg
All done in sab
W loop and smeared witch might have been a bad idea.
Bplus doing better w center inoc no smear.allothers smeared.
Incubate at 80 degree...pe6 only one no growth but has contam so idk....attachicon.gif20141202_125635.jpg

ad3b31e4cd21d56394216e811aff59d5.jpg

It appears that you have multiple germination points here but the arrows are where i would start. This is pretty basic and requires little more then gut feeling. I do however like to select as close to the division line that seperates the sectors. Its hard to tell because i dont know where all the germination was but it looks like the green arrow might be the fastest growing sector based on the size of it versus the surrounding sectors.

What i would do is to take 4 plugs and transfer to clean plates and let those 4 grow out half way. And monitor speed, sectoring, signs of rhizo or soon to be rhizo, density, and etc. Then take the rest of the plate and fruit it. You will not need to keep this as a master.

After the plates grow out, not all the way, select 1 plate and toss the rest into cold storage. This will be your working master, kind of anyway. The idea behind a master plate is that you want to minimize working with it so you dont risk contaminating it.

A master culture does not mean isolate or monoculture. An example would be plate 1-1 from my contaminated thread. I made 2 transfers from that plate and will not go back to it unless i loose all the progress. From the 1-1 plate i made 2 secondary masters which i will use to expand from there rather to continue isolation or to increase inoculant or to make another master. Following me? Its like making a copy of a document. I make 2 copies that i will use to make more copies if needed leaving the orginal document left alone but available if i ever loose the copies. That leads us to senescence. A copy of a copy of a copy will not be as clear or as dark as the original. So with that being said you want to minimize transfers so you dont want attempt isolating from any of these plates and should isolate from clone. Your first goal should be to get some nice agressive growth to feed grain to.

I hope this all made sense and i didnt confuse the crap out of you. I will be glad to answer any questions or give advice. I am no expert but can share my experience so far and what worked for me. The reality is at this point of the selection process is firing from the hip. Good luck buddy and keep me posted on this. PM at any time if you want also and that goes for anyone really. You guys/gals are my friends and enjoy hanging out here with all of you.
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#57 CatsAndBats

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Posted 02 December 2014 - 03:43 PM

Obviously I'm not doing agar work yet, but by next week when I am able to start, how would I approach it? I have two pieces of myc I was able to stab out of a valued fruit body in 4% dextrose..

 

just grow it out and clean it thru transfers?


Edited by catattack, 02 December 2014 - 03:44 PM.


#58 invisibilitysyndrome

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Posted 02 December 2014 - 05:05 PM

Ok so the reasoning behind smearing and was told by some in my town that smearing a spore across the plate alows fer a les concentrated amnt of spores and then less mating by thw end of the smear.
I totally undrrstand what you were saying up top no confusion and i will be working w one plate at a timw cold storage for the rest.
I did do a col grain on a plate i didn't take pic but its not doing very much....
I made 20 for fear of contam and in now way think i can keep up w all just gonna work w one or two. For thw time being

Edited by invisibilitysyndrome, 02 December 2014 - 05:07 PM.

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#59 Microbe

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Posted 02 December 2014 - 08:06 PM

Obviously I'm not doing agar work yet, but by next week when I am able to start, how would I approach it? I have two pieces of myc I was able to stab out of a valued fruit body in 4% dextrose..

just grow it out and clean it thru transfers?

place a single drop of LC on some agar and grow it out. Transfer as needed. Saw your P. Azz by the way. I cant wait to get into the wood lovers! Its going to be final project leading me into the world of edibles.
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#60 Microbe

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Posted 02 December 2014 - 08:13 PM

Ok so the reasoning behind smearing and was told by some in my town that smearing a spore across the plate alows fer a les concentrated amnt of spores and then less mating by thw end of the smear.
I totally undrrstand what you were saying up top no confusion and i will be working w one plate at a timw cold storage for the rest.
I did do a col grain on a plate i didn't take pic but its not doing very much....
I made 20 for fear of contam and in now way think i can keep up w all just gonna work w one or two. For thw time being

You are correct on the least amount of genetics you want to introduce into a plate. A single drop of spore solution or LC as worked for me. I use a loop when working from slant to plate only. But the smear is fine as long as you only touch the center of the plate once. Using a drop of liquid is more of a visual thing for me as i know its there while with a spore smear, it is mostly likely there but i always wonder if it really is. I wasnt saying using the loop was wrong. Everyone has their own preference. The only down side to smearing or scrape whatever its called is the hydration process of the spore. My spore syringes sit for a minimum of 48 hours before i disperse it on agar. Looking good so far.
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