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Mycobro420
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Clone on agar
#26744870 - 06/14/20 08:17 PM (3 years, 7 months ago) |
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How many transfers would a clone agar plate need to be clean enough for grain.an how many transfers for a single sector isolate from a clone
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A.k.a
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Clean is clean there’s no number of transfers. If you do the tissue sample well then zero transfers.
Isolates can take like 50+ transfers and seem to suck more often then not.
Usually with clones a couple transfers will be enough to get consistent results.
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Edited by A.k.a (06/14/20 09:30 PM)
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Mycobro420
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Re: Clone on agar [Re: A.k.a]
#26745791 - 06/15/20 06:58 AM (3 years, 7 months ago) |
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Ya I'm just worried about white mold an always thinks my cultures have them I ust to do fruit tissue peice to mea wraped in parafilm then sector the edges of dish to grain to new plate but I got trick on a few of them.so know I'm trying 2 transfers from master dish an see how that works I'm just confused what to look for cause I get rizomorphic growth from every dish including the flesh dish.also I have a flowhood
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Mycobro420
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How many transfers would you recommend I personally don't see much wrong with my petris no bateria spots no discoloration just white circular rizo growth.every now an then it wount be 100% circular Anni usually toss them
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mushpunx
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Can't answer that. A plate is clean when it's clean. It could take as a little as one transfer until it's clean.
You just want to make sure when you transfer that you take healthy tissue that is far away (and not on top of) bacteria and/or mold colonies. Bacteria can sometimes present as a thin film that can cover most of the top of the plate so watch out for that. Don't take tissue that isn't from the culture - white spots in random places on the plate could be one of two things, mycelium that fell off the wedge (maybe the wedge rolled a little), or they are mold colonies. It can be hard to distinguish between the two so I just don't transfer from them.
Don't get hung up on rhizomorphic and tomentose growth. A heavily tomentose (fluffy) culture will often turn rhizomorphic when transfered to media with less nutrients, and vice versa.
Just look for clean, healthy looking growth that's far from contams to transfer.
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Edited by mushpunx (06/15/20 07:53 AM)
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mushpunx
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Re: Clone on agar [Re: mushpunx]
#26745864 - 06/15/20 07:50 AM (3 years, 7 months ago) |
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Also dude you need to use more punctuation though, your posts are really hard to read. 
Again nobody can tell you how many transfers it will take to reach an isolate.
I don't think you understand what a sector is from the way you are using the term. Early transfers of a culture from spore or clone often have hundreds of strains present, so they will often look uniform and round. The more transfers you take though, there will be less and less strains present and you can start to see the lines of separation between individual sectors - there are simply too many strains present to see sectoring until a number of transfers have been made and many of the strains have been cut out.
When you have an isolated strain, it will be a monoculture - perfectly round with no visible sectors.
Clones will often have less strains present than multi spore cultures. Every once in a while you will take a clone and it will already be a monoculture (no sectoring). Sometimes you will be able to see sectoring right away and other times not. It usually takes some transfers to see obvious sectoring, but less transfers than an MS culture would take. I usually just keep clone cultures as is. Sometimes I will transfer them down to a monoculture. The most important thing is to label and test your clones , including potency.
With MS cultures it can often take 8 - 10 or more transfers until you see obvious sectoring. Then you want to take a transfer from each sector - the goal is to end up with many isolates from a culture at first, not one. Sometimes the transfers will continue to sector and you will need to continue to make transfers. Aim to end up with at least 10 isolated strains. You then label and store each isolate and fruit out each one and test them to compare. The best fruiting and most potent isolate is saved and put to slant.
Here is a really great thread about cleaning and strain isolation on agar, with great photos :
https://www.shroomery.org/forums/showflat.php/Number/18430998
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Edited by mushpunx (06/15/20 08:02 AM)
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Mycobro420
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Re: Clone on agar [Re: mushpunx]
#26746154 - 06/15/20 10:18 AM (3 years, 7 months ago) |
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Ok my bad I know my punctuation is sloppy bear with me lol.i understand that there are hundreds of strains on a single Petri sectoring is the fuzzy looking stuff in-between the rizomorphic growth. I'm not looking for a monoculture just good fruiting mycilum witch I know is more or trial and error. But really I'm trying to figure out if my agar is clean and free of hidden white mold.ill upload a pic as well mabey that can say more than I can.
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A.k.a
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Sectors are just different areas of growth, on a plate they’re like pizza slices. They can be rhizo or tomentose. There’s some good threads on it where guys outline sectoring on their plates.
This plate has some distinct ones.

Just below 9 to about 10 is a sector, and then there’s a super clear one with fuzzier myc from about 11-12
I did a quick rough edit to outline some of the sectors
 Even within those there are multiple smaller ones
Edited by A.k.a (06/15/20 10:37 AM)
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Mycobro420
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Re: Clone on agar [Re: A.k.a]
#26746292 - 06/15/20 11:11 AM (3 years, 7 months ago) |
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Ok having trouble uploading my picture let's see if this workshttps://www.shroomery.org/forums/files/20-25/224075427-IMG_20200615_120639400.jpg
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Mycobro420
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Ok I suck at this I don't know how to up load them
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Mycobro420
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Re: Clone on agar [Re: A.k.a]
#26746328 - 06/15/20 11:22 AM (3 years, 7 months ago) |
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With those pics I see how it's sectoring but another question how can u tell tomentose growth from white mold
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mushpunx
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Quote:
Mycobro420 said: Ok I suck at this I don't know how to up load them
Go to "pics" and click upload
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A.k.a
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Usually it looks slightly different than myc but it can be really hard to do.
Luckily I barely ever see mold that looks like myc. That I know of 
Most of the mold I get is dark or yellow, and if it is white it’s got long scraggly hairs.
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mushpunx
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Quote:
Mycobro420 said: With those pics I see how it's sectoring but another question how can u tell tomentose growth from white mold
When germinating spores we swipe them across the plate in a zig zag motion, back and forth. You should be able to see germination in that pattern and only taking tissue from that will increase your chances of taking clean myc.
When it is a clone or a transfer, be careful to take tissue that is expanding out from the wedge. Never colonies.
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mushpunx
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Quote:
Mycobro420 said: Ok my bad I know my punctuation is sloppy bear with me lol.i understand that there are hundreds of strains on a single Petri sectoring is the fuzzy looking stuff in-between the rizomorphic growth. I'm not looking for a monoculture just good fruiting mycilum witch I know is more or trial and error. But really I'm trying to figure out if my agar is clean and free of hidden white mold.ill upload a pic as well mabey that can say more than I can.
If all you are looking for is good fruiting myc then simple. Just keep taking clones and label them, then test them until you find a good one.
You should be able to get canopy flushes from MS grows. You need to keep improving sterile tek and how you dial in your tubs.
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Mycobro420
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Re: Clone on agar [Re: mushpunx]
#26746502 - 06/15/20 12:41 PM (3 years, 7 months ago) |
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I can get canopy flushes from Ms around 112g dry. I just ran my first bigger agar cycle to monotubs. I yeilded my best was 158g dry so it was a canopy sort to speak just looking for improvement cause I know 200g dry is possible. But why I made this thread is cause some of my bins caught trick on there first flush witch I know came from the agar or steril Tek or my g2g but first I wanted to figure out if the agar was the problem
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Mycobro420
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[embed=,]https://www.shroomery.org/forums/files/20-25/224075427-IMG_20200615_120639400.jpg[/embed]
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Mycobro420
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I still can't find how to post pic I got it posted on my account gallery but everytime I try to share it come up like the one before
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