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OfflineKmacmo
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Few questions about mycelium growth on agar pictures included
    #26392495 - 12/20/19 01:43 PM (4 years, 1 month ago)

hey everyone, was hoping some of you could answer these or provide some insight and knowledge to help me understand whats going on with these plates.

The green/yellow is on all of the plates with red food coloring, think the mycs sucking out the color. (not using food coloring anymore though)
The recipe for all of these is 10g LME (Brewing,sugar alternative)/10g agar agar vegan gelatin food grade for 500ml water.

Q1. Some of the mycelium around the transfer covers good area without being patchy or 'missing' area of myc.... while other areas are really patchy/spotty and mycelium is 'missing',
whats up with that, is it sectoring.

Q2.  Is translucent mycelium unhealthy/undesirable?  On nearly every plate theres translucent patches amongst the more white myc, its like it doesnt want to grow very much on those 'patches'..

Q3. If my tempratures are at a steady 15-20c wich is pretty cold, is it worth putting a heat mat down with a thick wire rack and a cardboard shoebox lined with some metal plates to disperse the heat (ive tested the contraption and the metal plates that would touch the petris is staying at a steady 22c all over with no 'hot spots') 1-2 inches over and just putting them ontop of that? My aim is not to 'heat' them too much just keep it at a steady 20-22c... it gets below 14c regularly in my house.

Here are some pics to go with it, not all at the same stage but just got onto T4 today (no pics of the T4's).

Any comments are much appreciated :smile:

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Edited by Kmacmo (12/20/19 01:45 PM)


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OfflineA.k.aM
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Re: Few questions about mycelium growth on agar pictures included [Re: Kmacmo]
    #26392630 - 12/20/19 03:14 PM (4 years, 1 month ago)

I’m new to agar but those look pretty sketchy.

What’s going on with 6-8??  It looks like a pic from the backside almost but the wedge is there. Never seen anything like that I’m curious what somebody who knows agar has to say.


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OfflineKmacmo
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Re: Few questions about mycelium growth on agar pictures included [Re: A.k.a]
    #26393440 - 12/21/19 03:07 AM (4 years, 1 month ago)

Yeah they all look sketchy too me, most of my mycelium seems to do this abit... But 6-8 are different wedges from different transfers cut with its own brand new sterile blade(I also flame steralize them).

I've had this happen before, that translucent stuff grows white after a while and does look alright.. Just gotta let those turn white and I'll see what's best to transfer (I hope) :P


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Offlinejcm4620
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Re: Few questions about mycelium growth on agar pictures included [Re: Kmacmo]
    #26393622 - 12/21/19 07:39 AM (4 years, 1 month ago)

id start over i dont think id trust any of those


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OfflineKmacmo
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Re: Few questions about mycelium growth on agar pictures included [Re: jcm4620]
    #26394085 - 12/21/19 02:09 PM (4 years, 1 month ago)

I don't know what's wrong but I have 10 or so newly inoculated plates with abit less nutrient in them so see how they go :/


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OfflineHighHarles
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Re: Few questions about mycelium growth on agar pictures included [Re: Kmacmo]
    #26394285 - 12/21/19 04:14 PM (4 years, 1 month ago)

Someone should correct me if I'm wrong but you shouldn't be using the tic tac toe method to be removing myc from plates for transfers. Research sectoring. You wanna cut out triangles of the furthest leading mycelium from the inoculation point. If you follow the ropey looking myc you'll see it has somethin like a main vein and branches out. You'll understand how genetics are specified a little bit better with a guided glance at some plates.


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OfflineKmacmo
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Re: Few questions about mycelium growth on agar pictures included [Re: HighHarles]
    #26394957 - 12/22/19 03:43 AM (4 years, 1 month ago)

I have researched quite a lot and looked at a lot of other photos of healthy myc... Was hoping to find out what's up with these just so I can learn something and not have it happen again ha.

I have inoculated another 10 clean plates, with 10g agar/7gram LME 500ml. Read that the mycelium should show more rhizomorphic growth because it will have to stretch and grow outwards to find nutes.

I can't compare my plates to anything I've seen online ha, so it's hard to see any 'sectors' on my plates. if all 20 of my new germination plates show the same fluffy shit and no rhizo/decent looking mycelium... I'll be stumped:/

Thanks for your advice


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Offlinejcm4620
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Re: Few questions about mycelium growth on agar pictures included [Re: Kmacmo]
    #26394966 - 12/22/19 04:13 AM (4 years, 1 month ago)

well first off if ppl knew what the species was you r tryin to grow would help a lot. second i wouldnt touch any of those with a 10 ft scalpel. looks like mold all over the place to me throw in a lil bacterial shit and you have yourself one hell of a science project goin on there. but u didnt describe your sterile tek at all sab???  flow hood ????  under the kitchen sink looks about rite lol  but anyways that kind of information is what ppl here need to have a better chance of helpin u correct your issue. as stated before id junk all of em and start over to b safe man cuz i dont think i c anything id even open up


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OfflineA.k.aM
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Re: Few questions about mycelium growth on agar pictures included [Re: jcm4620]
    #26395119 - 12/22/19 08:03 AM (4 years, 1 month ago)

I have a piece of tissue on a high nute plate that looks similar but not as extreme. It’s Burma and I’ve had nothing but mutants from them so far.


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OfflineKmacmo
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Re: Few questions about mycelium growth on agar pictures included [Re: jcm4620]
    #26395277 - 12/22/19 09:51 AM (4 years, 1 month ago)

Yeah I should have said its cubensis b+,  I use a SAB on my kitchen  counter top everything wiped/drenched in 70% isopropyl and then I spray disinfectant In the box and leave for atleast 15 mins with windows shut fans off.

I wash my hands/forearms and dry with clean towel, put on rubber gloves and drench hands/arms in iso... And everything that goes in gets a good wipe too.

Pull out my little inoculation loops pre sterile and flame it red it before sticking in the receiving dish and then carefully open my print and dab spores onto it, then swipe it around the plate 3 times. Job done.

Transfers done same way with pre sterile blades aslo flame steralized, one blade per transfer.

A.K.A the fresh plates I made are all 10g agar/7g lme so I'll see how they look different to the high nute plates :smile:

Thanks for the feedback has been very helpful guys :smile:


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Edited by Kmacmo (12/22/19 09:53 AM)


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Offlinejcm4620
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Re: Few questions about mycelium growth on agar pictures included [Re: Kmacmo]
    #26395338 - 12/22/19 10:19 AM (4 years, 1 month ago)

im willing to bet the issue is in your print maybe its totally fucked. and a new blade for every transfer i belive is totally a waste of blades and overkill by the way

get a new print and keep the same tek and c what happens


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Edited by jcm4620 (12/22/19 10:21 AM)


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OfflineHighHarles
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Re: Few questions about mycelium growth on agar pictures included [Re: Kmacmo]
    #26395349 - 12/22/19 10:24 AM (4 years, 1 month ago)

It may be benificial to stick to your sterile patterns, but you don't need  a new blade for every transfer. That's why there is flame sterilization. Do what makes you succesful, avoid wasting your resources n making more work than necessary.

keep isolating with your fingers crossed. Another option would be to just start fresh and acquire some new spores. Variety is lovely


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OfflineKmacmo
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Re: Few questions about mycelium growth on agar pictures included [Re: HighHarles]
    #26395425 - 12/22/19 11:09 AM (4 years, 1 month ago)

Yeah new spores, I'm UK and got these ones from zamnesia but their site has now removed all the mushroom stuff.

Have a new site in mind, not sure if it's a sponsor or not so won't mention the name but most reviews are good.

Also thinking of getting a kit I could just clone from that :laugh:

I just wanted to be super sure and minimalise any risk, don't want to do that for the future once I get it right... Just dip the blade in iso for a few secs and then flame steralize?


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Offlinejcm4620
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Re: Few questions about mycelium growth on agar pictures included [Re: Kmacmo]
    #26395430 - 12/22/19 11:16 AM (4 years, 1 month ago)

kits suck stay far far away from them if your already using agar you can pull off a grow on your own its not rocket science. just get new spores and stay with it man you will get there and when you do the reward will be much more worth it. kits r garbage and more than half of the time they are already contam when you get em  just hang in there


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Edited by jcm4620 (12/22/19 11:17 AM)


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OfflineKmacmo
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Re: Few questions about mycelium growth on agar pictures included [Re: jcm4620]
    #26395486 - 12/22/19 11:47 AM (4 years, 1 month ago)

I will continue the agar work its really enjoyable just need to get new spores :P
My first ever grow was from a kit wich gave me some nice big fruits I just wasnt into the agar at the time, should have cloned some of them.

Trying to cover all ground staying working with the spores and agar, the kit would ensure I get a fruit tho that sounds tempting :P

Heres some pics I took today of plates inoculated on the 5th december:

They look just like every other plate ive done, starts out with translucent 'myc' and then it will all whiten/fluffy up over time.


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Offlinejcm4620
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Re: Few questions about mycelium growth on agar pictures included [Re: Kmacmo]
    #26395505 - 12/22/19 11:57 AM (4 years, 1 month ago)

those dont look good


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OfflineKmacmo
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Re: Few questions about mycelium growth on agar pictures included [Re: jcm4620]
    #26395538 - 12/22/19 12:22 PM (4 years, 1 month ago)

its all growing directly from the spores (those black spot areas)

think the plates are abit dusty aswell :/


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OfflineRandalf the Grey
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Re: Few questions about mycelium growth on agar pictures included [Re: Kmacmo]
    #26395730 - 12/22/19 02:40 PM (4 years, 1 month ago)

:2cents:
Agar may be a bit stiff. Not that my way is THE way, but I can only speak from personal experience. 7.5g\500ml agar to water is my standard mix for MEA (15g malt extract)
Loop doesn't dig in to surface but it has some spongeyness to it. I think this helps the spores absorb some of the moisture from the agar and begin to germinate.
Currently reviving some old spores from both MSS and prints with this mix.
Also, I may be completely wrong on this part, but maybe try making an MSS with part of the print and give both methods a go.

Also, unless you were running low on agar or plates, I see no reason not to give it a go. One of the reasons I fell in love with agar is that even if a plate is contaminated, cut it transfer and try it again. You can do this almost ad infinitum until you get something usable. And I never expect my first plate from spores to go 2 grain. It is always going to go onto another plate.


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Randalf the Grey
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Edited by Randalf the Grey (12/22/19 03:38 PM)


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