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Offlinebigd17
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ms on agar, sectoring questions.
    #19468731 - 01/24/14 05:46 PM (10 years, 11 months ago)

So it's been 7 almost 8 days now since I innocculated 10 potato/honey agar dishes with Z-Strain MS solution and the babies are growing nicely. 

Not very big yet I'm figuring due to all of the various genetic material competing for nutes slowing things down, but I'm having an issue finding sectors.

All of the mycelium that have propagated all seem to be growing out in perfect circles and all of the information I have found on sectoring seems to be vague and ambiguous as to how to determine where the seperate sectors are. 

Judging by comparing mine to all of the pictures on here mine all look like monocultures, but I know this can't be true. 

Does anyone have any advice for me as I want to get my transferring done before these bitches hit the edges of the plates.


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"The secret to success is to make your Vocation a Vacation."
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Invisibleblindingleaf
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Re: ms on agar, sectoring questions. [Re: bigd17]
    #19468838 - 01/24/14 06:18 PM (10 years, 11 months ago)

ur first plate will likely not show sectoring (because of what u said, many many strains).  transfer from anywhere if it all looks the same, or whatever leading edge u may see.  sectoring should be apparent hopefully next plate or two.


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InvisibleEnigma1
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Re: ms on agar, sectoring questions. [Re: blindingleaf]
    #19468863 - 01/24/14 06:26 PM (10 years, 11 months ago)

Remember transfer no larger than a grain of rice.It took me 3 transfers from a clone to get a mono.

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OfflineRogerRabbitM
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Re: ms on agar, sectoring questions. [Re: Enigma1]
    #19468907 - 01/24/14 06:36 PM (10 years, 11 months ago)

You also better isolate several strains.  Not all are going to be good performers.  It usually takes two to four transfers before you start seeing sectoring unless you're really good at swiping very few spores.
RR


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Offlinebigd17
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Re: ms on agar, sectoring questions. [Re: RogerRabbit]
    #19469764 - 01/24/14 10:13 PM (10 years, 11 months ago)

Yeah I dropped them straight from the syringe onto the agar there was no swiping lol... is that what I should do in the future?

This would probably explain why I seem to have different spote of mycelium forming all over the place...  Whoops.




That's my still air box they are in, I will post some closer pics up once they've grown out some and I pull them out to work on them.


--------------------
"The secret to success is to make your Vocation a Vacation."
      -Mark Twain

Confucius say,"man who stand on toilet, high on pot."

The answer to the question never asked is always no.

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Re: ms on agar, sectoring questions. [Re: bigd17]
    #19470803 - 01/25/14 06:50 AM (10 years, 11 months ago)

if i use a syringe, i just do one tiny tiny drop after rapidly shaking the syringe both before flaming (outside the SAB) and right before i plunge the tiny drop out (in the SAB).  the shaking disperses the spores (in case there are not that many in the syringe to begin with) and the tiny tiny drop (hopefully) makes it so there are not so many deposited.  the fewer that are deposited, the better (if u wanna see sectoring vis a vis less mating, less strains/sectors forming).  thats what RR meant (I'm pretty sure) when he said "unless ur really good at swiping very few spores"


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A few thoughts on cultivation
MICROBIAL HUSBANDRY!!!!

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Offlinebigd17
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Re: ms on agar, sectoring questions. [Re: blindingleaf]
    #19471670 - 01/25/14 11:51 AM (10 years, 11 months ago)

Ah ok I thought he was talking about using a sterile cotton swab lol.  So I know those pics aren't very good but based on how big the myc looks compared to the dish do I need to wait longer before I bother trying to sector?


--------------------
"The secret to success is to make your Vocation a Vacation."
      -Mark Twain

Confucius say,"man who stand on toilet, high on pot."

The answer to the question never asked is always no.

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Re: ms on agar, sectoring questions. [Re: bigd17]
    #19472839 - 01/25/14 04:22 PM (10 years, 11 months ago)

just don't let it get to the edge of the plate.  i will look at my plates after i wake up/drink coffee.  if there is enough room for it to grow that it will not touch the edge while I'm at work, i will wait to transfer (if I'm looking for sectoring/thicker growth) till i get home.  if it looks like it will touch the edge while I'm away, but i still do not see sectoring, i will take from the edge that is furtherest along, or whatever part of the mycs perimeter looks thickest.  since u r on ur first plates, it may take 2-4 more transfers to see sectoring, so don't worry.  sometimes u get sectoring on second plate, sometimes on 5th.


--------------------
A few thoughts on cultivation
MICROBIAL HUSBANDRY!!!!

The whole is greater than the sum of its parts

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Offlinehushmush

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Re: ms on agar, sectoring questions. [Re: bigd17]
    #19472881 - 01/25/14 04:34 PM (10 years, 11 months ago)


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Re: ms on agar, sectoring questions. [Re: hushmush]
    #19472902 - 01/25/14 04:40 PM (10 years, 11 months ago)

:whathesaid:
yea i love that thread


--------------------
A few thoughts on cultivation
MICROBIAL HUSBANDRY!!!!

The whole is greater than the sum of its parts

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Offlinebigd17
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Re: ms on agar, sectoring questions. [Re: blindingleaf]
    #19474779 - 01/26/14 12:18 AM (10 years, 11 months ago)

Oh yeah I definitely have that thread bookmarked already haha I do as much research as possible before I ask questions.

What I've realized is that instead of noc-ing 10 plates and saving 6 I should have noc-ed like 2 plates and saved the other 14 for transfers...:facepalm3:  I really wasn't expecting to have 100% success rate on my first try esp with just a GB and a half ass pressure cooker

But it's ok I have 2 more sleeves of dishes before I'll need to get more.  I will do what I can with what I have.

QUESTION:  I have a 23 qt PC on the way from Amazon, would it be crazy to sterilize some jars of grain and just throw the whole of the weak/seemingly slow moving cultures into there? (so as to not waste time transferring them and potentially still get something out of them?) Or is that just a stupid idea?


--------------------
"The secret to success is to make your Vocation a Vacation."
      -Mark Twain

Confucius say,"man who stand on toilet, high on pot."

The answer to the question never asked is always no.

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Invisibleblindingleaf
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Re: ms on agar, sectoring questions. [Re: bigd17]
    #19475499 - 01/26/14 06:48 AM (10 years, 11 months ago)

not sure what u mean. 
ur grain jars have growth, but it is slow, so u wanna re- pc the jars and then re use them by inoculating with additional wedges?  I'm assuming it can be done.  i have done that with jars that showed no growth from MS, but not agar, so i guess it would also happen with agar growth.  be careful that the grains are not too dry though


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A few thoughts on cultivation
MICROBIAL HUSBANDRY!!!!

The whole is greater than the sum of its parts

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Offline1down5up
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Re: ms on agar, sectoring questions. [Re: blindingleaf]
    #19475661 - 01/26/14 07:57 AM (10 years, 11 months ago)

If you knock up your grains with myc from your first plates, the performance won't be as predictable as it would wehn you get closer to a tested monoculture, but you can still do it.  I took myc from a clone after first transfer and knocked up some WBS.  They seem to be doing good :thumbup:

Good luck brother!


--------------------
~~Everything is relative~~


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Offlinebigd17
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Re: ms on agar, sectoring questions. [Re: 1down5up]
    #19476045 - 01/26/14 10:17 AM (10 years, 11 months ago)

Yeah that's what I mean, using the first cultures from MS onto agar and putting them into grain, just because I have too many to work with reasonably but I don't want to waste them since I took the time to make them and let them grow a bit.

I see what you mean about predictable growth but that's why I will be selecting the best 2 or 3 plates to take sections from for transfers and use the other ones to just throw into grain rather than attempting to store them.

Theoretically, if I had a quart jar of rye berries and took a silver dollar-size culture and cut it into 10-15 small pieces and threw them into a jar then it should help to get it to colonize quicker even if they are unpredictable and slow?


--------------------
"The secret to success is to make your Vocation a Vacation."
      -Mark Twain

Confucius say,"man who stand on toilet, high on pot."

The answer to the question never asked is always no.

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Offline1down5up
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Re: ms on agar, sectoring questions. [Re: bigd17]
    #19476343 - 01/26/14 11:41 AM (10 years, 11 months ago)

I don't really see anything wrong with experimentation in this hobby.  Some of the more practiced growers may tell you that 1st transfer might be a waste of time, because it isn't much different from doing MS, but they have the benefit of experience over people like me....but you have to get experience somewhere man....so if you wanna use it, giver. 

Quote:

bigd17 said:
I see what you mean about predictable growth but that's why I will be selecting the best 2 or 3 plates to take sections from for transfers and use the other ones to just throw into grain rather than attempting to store them.





Even taking your two best, you still won't get anything more predictable or more random than if you used MS...but that doesn't mean you shouldn't try....you'll still get mushies if you grow'em out right :thumbup:


--------------------
~~Everything is relative~~


A Simplified Overview of Mushroom Cultivation Strategies  -  RR says  -  EvilMushroom666's Take on BRF Cakes  -  Frank's list of goodies  -  Cronicr's Goodies


No one is placed higher than another no matter race or creed or gender, we must teach forgiveness and compassion for all life.  J.L.

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Offlinebigd17
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Re: ms on agar, sectoring questions. [Re: 1down5up]
    #19476644 - 01/26/14 12:44 PM (10 years, 11 months ago)

Hell yeah thanks for the encouragement!

and I can see how it is a waste of time, but since their already on the dishes and there's not much else I can do with them aside from store them in a fridge for a couple months, I figure I might as well use them.


--------------------
"The secret to success is to make your Vocation a Vacation."
      -Mark Twain

Confucius say,"man who stand on toilet, high on pot."

The answer to the question never asked is always no.

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Re: ms on agar, sectoring questions. [Re: bigd17]
    #19477107 - 01/26/14 02:29 PM (10 years, 11 months ago)

if u do print/syringe to first plate and then transfer, at that point the only benefit is knowing if it was clean or not.  i still would transfer once to 2nd plate and then use a wedge, just in case.  from that second plate, use the leading edge, most thick/rhizo growth.  i use a triangle the size of half of a quarter.  i'd cut it into two.  seems like taking the size of a half dollar and cutting it into 10-15 pieces would be a lot of work, and u'd have to keep petri open a while, giving con tams a longer window of opportunity.

and yea like 1 down said, use them on grains to start a grow.  u don't have to isolate to grow, and it will give u practice setting up conditions for growing, seeing how ur environment is, and helping u decide if u wanna do mono, trays, etc.


--------------------
A few thoughts on cultivation
MICROBIAL HUSBANDRY!!!!

The whole is greater than the sum of its parts

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