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Re: AGAR ENVY! (Anything and All things agar!) [Re: nmd_myco] * 1
    #26989922 -

nmd_myco said:
why wouldn't i just drop a slide cover on every plate from MSS?

drop, cover, wait? seems like a great way to get a good start from spores... maybe wait till you see germination and then drop the glass as a matter of course.


Hi nmd_myco ... yes, if that tek works I will use it every time for new cultures. Also for clones it would be good, especially small mushrooms like pans where it is harder to get a clean piece.

It is very simple to do. And what also catches my great attention is that in the original research paper they say they found out, that the resulting culture was showing better growth than the parent culture. Their theory was, that the fight through the barrier was only possible for the most healthy mycelium.

However, I will need to see through more transfers and fruiting to see if it really works.

But so far the results look promising. Maybe more people will try it. It could be a nice adding to the known agar teks.


A.k.a said:
Wow I’m surprised you were able to get that wedge into the new plate without showering spores all over.

Most of the time when I take wedges from plates with tons of mold I get a bunch of satellite colonies on the new plate.


Thank you very much A.k.a !

I think the contaminant is mostly bacteria, but it messes with the mycelium and makes it act weird in fear for its life. At least that is what I hope. If it is mold I might have isolated the best growing mold from the parent plate. I think I will have an update in a week ... going to do transfers tomorrow.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: nogoodnamesleft]
    #26989969 -

nogoodnamesleft said:
nmd_myco said:
My germ plates are a mess. Was thinking about doing the same thing when my grain gets here.  The timing should be right.



But like...that’s not what the TEKs say to do... LOL

D3monic said:
I wouldn’t send any of those personally. But I’m picky. I don’t even feel these are ready


Y’all on this site are some of the most uppity people...

Let’s just step back and appreciate some progress shots and some future plans. Also please note that there are 3 of 75 plates pictured...

No agar to grain right now.

I do have a few zones that will be getting transferred...



Lol, whaaaat? No one is being uppity. We are just informing you that none of those plates look clean enough to send like you had mentioned. For what it’s worth I thought the black plates agar was pretty.

The last plate pictured, maybe it’s a bad pic but looks like nothing but bacteria to me. Maybe drop the ego down a few notches and learn to take constructive criticism. No one wants to see you waste your time. I could never imagine sending a swab plate.. rarely do I ever even send a plate that is only on T2. It would have to look immaculate.


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D3’s Glog
D3's Agar TEK
Agar Porn Gallery
RW x MelMak Grow Along Thread

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Re: AGAR ENVY! (Anything and All things agar!) [Re: D3_Myc]
    #26990006 -

Just a lil FYI, I did a comparison on the difference Charcoal made to my agar. I did not like the amount of sediment I got (obviously) but the results were pretty good. I tested both a batch of plates and a whole heap of no pours, and did one Half Strength MEA and one Charcoal ME (2 grams of charcoal per litre).
I transferred nearly every variety I have one to one, no donor sample being below T2 I think, so while this isnt a proper monoculture test, it did give me a birds eye view on whether adding charcoal (I specifically used bone charcoal) was beneficial or not.

The difference in whether it was better for the myc in terms of grow speed was kind of iffy in some cases but I tallied up which one of the two samples was the "better looking" one in terms of both growth speed and uniformity.

Without Charcoal (.5 strength MEA) , 7 of the No-Pours and 3 of the Petri plates looked better.
3 of these samples contaminated, probably due to sterile tek.

With Charcoal (2 grams for every 10 grams of Malt), 14 of the No-Pours and 2 of the Petri plates looked better.
5 of these samples contaminated, probably due to sterile tek.

I have no idea if this is useful to anyone but my take from this run was that the charcoal wasnt really worth it despite showing slightly stronger results.


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Re: AGAR ENVY! (Anything and All things agar!) [Re: ManifoldPrime]
    #26990054 -

Yea, I don’t see any added benefits of the charcoal over not other than color contrast. Black does look sexy though. Hoe fine was your powder?  The one I use is made from bamboo I believe and very fine.


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D3’s Glog
D3's Agar TEK
Agar Porn Gallery
RW x MelMak Grow Along Thread

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Re: AGAR ENVY! (Anything and All things agar!) [Re: D3_Myc]
    #26990083 -

@xspak:
Very interesting! Did you sterilize the slide (if so, how) or just took one from a fresh pack?

Just did the poor man's version of the microscopy slide:

-Sterilize scalpel and cut a small well in a new agar dish (a bit off center)
-take a small transfer from the bacterial plate and put it in the well of the new plate
-sterilize scalpel again
-cut a large square agar wedge from the new plate and put it over the well to cover the transferred agar piece.

Will post results as soon as available.



Edited by sporecap (10/17/20 02:16 PM)

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Re: AGAR ENVY! (Anything and All things agar!) [Re: sporecap]
    #26990186 -

Hello sporecap,

thank you for your interest in the tek !


I took normal non-sterile slides 22 x 22 mm size - they are about 5 to 8 €.

I wrapped them individually in aluminum foil and then sealed them individually in sterilizer pouches, like these:

https://smelltest.eu/en/product/self-seal-sterilization-pouches/


These are used by dentists and tattoo folk to sterilize their stuff and keep it sterile afterwards, just like syringe needle packing they have one side foil and the other side some sort of paper or tyvek or so. they keep the goods sterile for a long time. On a side note: the larger ones of these are great to sterilize glass petri dishes and keep them sterile until you use them.

I made a whole bunch of them and put them into a jar that was not tightly sealed and sterilized them in the pressure cooker together with my agar.

The sterilizer pouches are not really necessary, I just use them because they are not expensive and I'm paranoid about contamination.

You can just wrap the slides in aluminum foil individually and then wrap them together in foil and/or put them in a jar and sterilize.

You maybe even can sterilize them in an oven.


To put them into the petri dish I used forceps that I sterilized in the flame just like a scalpel. Better are thin forceps I think so they glow quickly.


Your tek will also work I think, it is very similar to what I read here:

https://www.researchgate.net/publication/248894057_A_simple_and_effective_method_for_the_elimination_of_bacteria_from_fungal_cultures

If both these teks work, your way would be simpler because no need for the cover slide.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: xspak]
    #26990204 -

I use those bags for sets of scalper/scissors/tweezers. i have two sets. every time i run the PC, i throw any dirty sets in. each agar session, i bust out a clean set. I would put a loop in too, but I am currently using disposable loops. i may test one in its own pouch to see if it melts.

I like the idea of having a few of these plate covers individually wrapped and read to go in the sterile packaging. I have 3 sizes of those bags and so far have only found a use for the medium size which perfectly fit the tree tools i keep in sets. I need to do a bit of research about the sterile bags and how good they are at staying sterile in the bag and for how long.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: nmd_myco]
    #26990207 -

I tried those pouches but my contents got soaking wet. Maybe it was a bad brand


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

D3’s Glog
D3's Agar TEK
Agar Porn Gallery
RW x MelMak Grow Along Thread

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Re: AGAR ENVY! (Anything and All things agar!) [Re: D3_Myc]
    #26990233 -

nmd_myco said:
I use those bags for sets of scalper/scissors/tweezers. i have two sets. every time i run the PC, i throw any dirty sets in. each agar session, i bust out a clean set. I would put a loop in too, but I am currently using disposable loops. i may test one in its own pouch to see if it melts.

I like the idea of having a few of these plate covers individually wrapped and read to go in the sterile packaging. I have 3 sizes of those bags and so far have only found a use for the medium size which perfectly fit the tree tools i keep in sets. I need to do a bit of research about the sterile bags and how good they are at staying sterile in the bag and for how long.


I do it the same way with my forceps and scalpel every agar or grain run. I still use the flame though, but to give them a nice basic cleaning. I guess aluminum foil works just the same for all our purposes.

Most of these bags have that autoclav indicator that changes color when it reached a certain temperature. So at least we know that the pressure cycle went up to the desired temperature.

Normally from the specification of these bags they should be good for many months. Just like syringe needles and all the other medical stuff they pack in them.

The sterile swaps I use to put spores on a plate are also wrapped in these my the manufacturer (too cheap to make them yourself unless the budget is suuper tight).

D3monic said:
I tried those pouches but my contents got soaking wet. Maybe it was a bad brand


Hi D3monic !

I think the pouch brand was fine. But in our cases, I believe we still need to wrap them in aluminum foil or put them in a jar or best even both. The medical autoclaves have a dry cycle where they make everything in the autoclave bone dry with a vaccum or so. Our pressure cookers don't do that. I have not tested to use the pouches without wrapping them in foil - but I guess it might lead to them getting very wet.

So in the end these pouches can be of some use but because we still need to wrap them in foil or put in jars etc. they are not a huge advantage. The foil alone will do the job and once the idems wrapped in foil are cooled and kept in for example a zip-bag (leaving them wrapped in the aluminum foil, too ) - that is just as good as these pouches. Which may be that they are not mentioned on the shroomery very often.

They are a nice gadget and I like the temperature indicators many of them have. But certainly indeed they are not needed to grow mushrooms.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: xspak]
    #26991089 -

Hi all...What an amazing thread for a baby agar freak!! I have been contentedly transferring from dish to dish and learning as I go but I'm a bit confused by this batch. These are photos taken of dishes containing the first transfers from a mazatec culture that was germed from a syringe.  The inoculation from the syringe was on 9/26 and the dish developed very slowly compared to other plates from other syringes.  Upon finally seeing potential mycelial development, I took samples and transferred them to these dishes on 10/13.  My concern stems from being unsure about the health of the original germed culture and how wispy, disorganized and totally diffuse the developing cultures are looking.  They look more like the iris of an eye than other plates and I don't know enough to make the call on whether I'm growing out mycelium colonies or some rough mold/bacterial beast that is slouching hard toward Bethlehem.  In particular, this looks like some of Bod's shots of cobweb mold in Tmethyl's thread.  OTOH, despite the internal chaos, the development looks nice and  circular - a victory one way or the other!!  In sum, the overall circle of growth looks very similar but the disorganized chaos within  the circle looks like what I imagine the cosmos looked like after the Big Bang.

(I apologize in advance for the photography.  Due to using PP5 dishes, the wispy diffusion of the images, and my current lack of mycelial photographic chops, I fucked around with contrast, etc., so each image has a photoshopped doppleganger which -in my mind- helps to better demonstrate the structure of whatever it is that I'm growing than the original shots.  Of course, your mileage may vary!)

I'm hoping to get your thoughts regarding the viability of these plates. I'm not so much concerned about the details in each dish.  It's kind of a yes/no proposition.  If primarily mycelial, I will continue to work with it and clean as necessary. If I'm germinating universes of contamination, it's off to the dish disposal.  One way or the other, I'm learning a shit-ton.  Or perhaps a fuck-ton.  Thanks much, in advance!! 

1A


1B


2A


2B


3A


3B

Edited by mayan (10/18/20 08:18 AM)

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Re: AGAR ENVY! (Anything and All things agar!) [Re: mayan]
    #26991141 -

Yea it’s so hard to tell with the frosted pp5 dishes. I bought a bunch and stopped using them myself as the clarity was too hard to discern what was going on. The highly contrasted dish looks like it’s probably just tomentos growth but age of plates is a bit concerning. They should of grown in several times over already. Can you give a bit more background, average temp where you store your plates and agar recipe.


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

D3’s Glog
D3's Agar TEK
Agar Porn Gallery
RW x MelMak Grow Along Thread

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Re: AGAR ENVY! (Anything and All things agar!) [Re: D3_Myc]
    #26991183 -

Thank so much for your quick response, D3 (and thanks for your amazingly beautiful, informative and inspiring Agar thread).  The plates in the photos were started on October 13. It's interesting, I was actually concerned that these started growing quickly after the transfer - there was a discernable nimbus/halo of growth away from the pucks within 24 hours the transfer to the plates - thus fueling my always-active newbie paranoia that this was a mold of some sort. 

The average temp in the room is in the 70-73F range. The original syringe was commercial, multi-spore.  I tried to germ three plates from the same syringe but this was the only one that produced.  (I seem to have more germination success from print than syringe but the other dishes failure to do anything made me wonder about the overall viability of this particular syringe.  Of course, it could be my syringe germination technique that needs tweaking!)

The agar recipe was essentially - 6g of BRF, 9g of agar and 400 ml of water.  No coloration added. 

Oh!  And I hear you about the PP5s. Just getting started, it's nice to have re-usable dishes because of my learning curve.  As well, I'm trying to be sustainable and reducing my footprint for posterity. Of course, OTOH, just getting started, it would be nice to be able to see what I'm doing doing without peering through layers of mist and condensation - particularly with dealing with the further murk of a plastic tote SAB.  I've found that using a Carson magnifying lamp to be very helpful, however.  As well, I've been configuring adjunct lighting for the SAB and that has been paying dividends. 

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Re: AGAR ENVY! (Anything and All things agar!) [Re: mayan]
    #26991261 -

Oh sorry, was half asleep when I responded. Was thinking the xfers were in September. You’re right in suspecting the rapid growth. They seem pretty developed for having been xferred on the 13th. Low nute and aggressive myc could be a possibility but let’s air on the side of caution. The good news is you’re only waisting time and a small amount of agar mix if you do xfer some more.

Here’s a few plate comparisons I just snapped in the lab. Obviously different strains and agar recipes so grain of salt and all.

This is my slowest growing strain. I take xfers late at night so can almost consider this being the 15th more than 14th




Where this is fairly typical growth speed. Taken on the 11th



And this is one my more aggressive strains taken on the 8th.



Comparing the growth speeds of those to the plates you shared. I’d say yours are progressing alarmingly fast. What you could do is take xfers and then watch the donor plates and see what they do as they progress. Or even take a xfer in the safety of your sab then sacrifice the donor plate by bringing it out in the open, taking the lid off and giving it a good inspection. That way your transfers are already done and you can see/take pics of what’s actually going on on that plate.

If that makes sense?


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D3’s Glog
D3's Agar TEK
Agar Porn Gallery
RW x MelMak Grow Along Thread

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Re: AGAR ENVY! (Anything and All things agar!) [Re: D3_Myc]
    #26991383 -

Makes a lot of sense.  I'm tic-tac-toeing some other plates into oats today; I gotta do the sanitization/sterilization dance, so why not? This is helpful - thanks!  I do not have a good feeling about these - I looked at the plate with higher magnification and these's no flow - it's all pixilated, so I'm about ready to make the call at any rate. If so, garbage disposal - where, I hope, albino cubnesis mutants get eaten by the albino alligators in the sewer system.

Edited by mayan (10/18/20 12:26 PM)

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Re: AGAR ENVY! (Anything and All things agar!) [Re: mayan]
    #26993541 -

I put this in the fridge and checked on it after a few weeks. The whole thing was frozen. It's been sitting out for a 4 days and looks like not all of it died. Kinda cool in the sense that it was unintentional isolation. Do you think this is any indicator that the myc that survived did so because it's healthier/stronger?



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Re: AGAR ENVY! (Anything and All things agar!) [Re: Doctor Mario]
    #26993644 -

please explain "unintentional isolation"

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Re: AGAR ENVY! (Anything and All things agar!) [Re: nmd_myco]
    #26993666 -

nmd_myco said:
please explain "unintentional isolation"


When we transfer small pieces of myc to a new plate, we're isolating the culture to an extent. When the mycelium froze (unintentionally) some of the culture lived thus isolating itself from the rest of the dead culture. Notice how the new growth is a lot more rhizomorphic than the area it grew off of? I just wonder if the myc that lived was stronger/healthier than the myc that died.

Theres lot of good sections to transfer from and this is all theoretical with nothing to back up the theory but what if freezing the culture killed the weaker genetics? That would mean that isolating the best candidate of the culture just got easier, right?


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Edited by Doctor Mario (10/19/20 09:06 PM)

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Doctor Mario]
    #26993759 -

Did some transfers today.  1 transfer to LC, another transfer to a new dish.  Any input would be appreciated!

I let the GT and RW go a little longer than i meant to.  I was busy and couldnt get to them for a couple days.

GT Transfer 2


PE6 first transfer


RW Transfer 2


I use long tweezers, like you wuould get in a small tool kit for transfers so the long skinny marks are me cooling the red hot tweezers.

Edited by maniac1886 (10/19/20 10:12 PM)

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Re: AGAR ENVY! (Anything and All things agar!) [Re: maniac1886]
    #26993799 -

I have a friend. (Not close but we both grow) who regularly freezes spawn jars at 100%


--------------------
"Remember, kids, the difference between science and screwing around is writing it down" -adam savage
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Learn the tried and true way to grow mushrooms
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Re: AGAR ENVY! (Anything and All things agar!) [Re: tryptkaloids]
    #26993829 -

Is there any other purpose for freezing spawn jars other than to stall the growth?


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