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OfflineFert Nitty
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Question about multispores
    #19363813 - 01/03/14 03:07 PM (10 years, 27 days ago)

If i took a sterile print from a large healthy Cubensi fruit, is that still considered multispore inoculation when I knocked up my PF jars? Or is it different because it all came from a strong dominant singular mushroom? I'm just trying to get my terminology right and make sure I'm doing everything as best as possible. I also have agar plates and was thinking about starting a culture using a spore syringe I made from a print.


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Re: Question about multispores [Re: Fert Nitty]
    #19363825 - 01/03/14 03:10 PM (10 years, 27 days ago)

yes multi spore is any time you use spores, a single print will create hundreds if not thousands of "strains" when used


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OfflineFert Nitty
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Re: Question about multispores [Re: cronicr]
    #19363852 - 01/03/14 03:15 PM (10 years, 27 days ago)

I have been knocking up my last few batches with syringes ive made, and they are much faster to colonize and the rhizo mycelium is WAY more apparent than the syringes i bought from a vendor and inoculated at the same time. would it be beneficial to use these spores on agar and and then transfer the established rhizo mycelium to another plate? this is my first attempts with agar.. thank you for any input


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Re: Question about multispores [Re: Fert Nitty]
    #19363867 - 01/03/14 03:18 PM (10 years, 27 days ago)



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OfflineKnownPown
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Re: Question about multispores [Re: cronicr]
    #19364007 - 01/03/14 03:45 PM (10 years, 27 days ago)

The spores you made into syringes are probably colonizing faster due to being fresher, or you just got a good set of genes!

Once you get some spores or live myc to agar, you can let it grow out and isolate the sectors like the guides cronicr posted =) Goodluck man!


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Re: Question about multispores [Re: KnownPown]
    #19364024 - 01/03/14 03:49 PM (10 years, 27 days ago)

yup, thanks! im gonna clean my house toss all that in mt SAB and knock em out tonight! I got a syringe from freespores.com and i think it is bunk because its been 2 weeks since i inoculated 6 jars and all but 2 got contaminated and those ones show absolutely no growth, the ones i did myself with my own spores are almost finished colonizing...


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Re: Question about multispores [Re: Fert Nitty]
    #19364152 - 01/03/14 04:21 PM (10 years, 27 days ago)

awesome stuff man! keep us posted


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OfflineFert Nitty
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Re: Question about multispores [Re: cronicr]
    #19364169 - 01/03/14 04:25 PM (10 years, 27 days ago)

I will keep you posted. so I should put a couple ccs from my syringe on the middle of my plates using a glove box and I can incubate them with my pf jars, right?


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Re: Question about multispores [Re: Fert Nitty]
    #19364177 - 01/03/14 04:26 PM (10 years, 27 days ago)

just a drop or two :thumbup: and use a sab there much more effective


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Re: Question about multispores [Re: cronicr]
    #19364232 - 01/03/14 04:36 PM (10 years, 27 days ago)

i have a SAB, i just checked out my 'prepoured' agar plates and one of them has a ring of black all the way around the outside of it, im assuming contamination?


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Re: Question about multispores [Re: Fert Nitty]
    #19364241 - 01/03/14 04:37 PM (10 years, 27 days ago)

probly, and:thumbup: on the sab i'm not a fan of glove boxes


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OfflineFert Nitty
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Re: Question about multispores [Re: cronicr]
    #19364256 - 01/03/14 04:41 PM (10 years, 27 days ago)



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Re: Question about multispores [Re: cronicr]
    #19364270 - 01/03/14 04:44 PM (10 years, 27 days ago)

Quote:

cronicr said:
probly, and:thumbup: on the sab i'm not a fan of glove boxes



just curious but why don't you like glove box SABs, mine doesn't have full gloves but id dose have the sleeves from a tyvek paint suit. is that ok or should I remove them?


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Re: Question about multispores [Re: MudaFuka]
    #19364278 - 01/03/14 04:46 PM (10 years, 27 days ago)

much easier to flame stuff when the gloves aren't attached:thumbup:


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OfflineFert Nitty
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Re: Question about multispores [Re: cronicr]
    #19364329 - 01/03/14 04:54 PM (10 years, 27 days ago)

well i got a glove box and some arm long gloves, but i usually dont use gloves


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Re: Question about multispores [Re: Fert Nitty]
    #19364456 - 01/03/14 05:12 PM (10 years, 27 days ago)

a glove box is basicly just a sealed sab, if your is just 2 holes for your arms thats a sab


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Re: Question about multispores [Re: cronicr]
    #19364475 - 01/03/14 05:15 PM (10 years, 27 days ago)

i can make it sealed with gloves on it, so it doubles as both if need be


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Re: Question about multispores [Re: Fert Nitty]
    #19364490 - 01/03/14 05:17 PM (10 years, 27 days ago)

well in this hobby it won't need be:thumbup:


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Re: Question about multispores [Re: cronicr]
    #19365988 - 01/03/14 09:47 PM (10 years, 27 days ago)

is there a decent way to take rhizomorphic mycelium from a colonized pf cake?


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Re: Question about multispores [Re: Fert Nitty]
    #19366496 - 01/04/14 12:20 AM (10 years, 27 days ago)

take it to the sab and take a piece from the middle,use a little less nutes in your agar to manipulate the rhizzo growth but generally they tend to take right off on agar anyway as brf is alot more nutritious then the agar:cool:


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Re: Question about multispores [Re: cronicr]
    #19366542 - 01/04/14 12:46 AM (10 years, 27 days ago)

what is so significant about rhizo growth of mycelium?
I had a search done and only results I got was that mycelium grows like that because of being in search of more nutrients and that its a bad sign??
my first set of jars inoculated with mexican cubes had normal growth and the new ones that i have with PES amazonian have the rhizo growth


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Re: Question about multispores [Re: bolbol]
    #19366551 - 01/04/14 12:53 AM (10 years, 27 days ago)

not a bad sign lol, it's just easier to see what your working with on plates, it's a bad sign if you stick a grain on cardboard where it goesright rhizzo because theres no food for it to eat or at least very minimal, in things like grain jars it's:thumbup:


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Re: Question about multispores [Re: cronicr]
    #19369646 - 01/04/14 06:27 PM (10 years, 26 days ago)

Quote:

cronicr said:
take it to the sab and take a piece from the middle,use a little less nutes in your agar to manipulate the rhizzo growth but generally they tend to take right off on agar anyway as brf is alot more nutritious then the agar:cool:





my agar plates are prepoured, next i will be making my own.


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Re: Question about multispores [Re: Fert Nitty]
    #19369649 - 01/04/14 06:28 PM (10 years, 26 days ago)

no worries at all you will still see good growth


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Re: Question about multispores [Re: cronicr]
    #19400731 - 01/10/14 07:17 PM (10 years, 20 days ago)

well i just got over my cold, so i just dropped a couple cc's out of my syringe into an agar plate in my s.a.b. so, just sticking it in my incubator should be the correct conditions for my culture?


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Re: Question about multispores [Re: Fert Nitty]
    #19400753 - 01/10/14 07:24 PM (10 years, 20 days ago)

YEP


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Re: Question about multispores [Re: cronicr]
    #19448198 - 01/20/14 03:17 PM (10 years, 10 days ago)

I just today spotted the first mycelial growth on my agar plate... I was beginning to think it was a dud. joy!


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Re: Question about multispores [Re: Fert Nitty]
    #19448203 - 01/20/14 03:17 PM (10 years, 10 days ago)

it is kind if fuzzy like mycelium lacking FAE.


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Re: Question about multispores [Re: Fert Nitty]
    #19448567 - 01/20/14 04:47 PM (10 years, 10 days ago)

they don't need fae at that point:thumbup:


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Re: Question about multispores [Re: cronicr]
    #19448831 - 01/20/14 05:44 PM (10 years, 10 days ago)

oh i know, i was just describing the way it looked, like it was stretching out hard. it should grow fast from here now,right?


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Re: Question about multispores [Re: Fert Nitty]
    #19508729 - 02/01/14 11:33 PM (9 years, 11 months ago)

I realize this isn't ideal, but finally this is the growth I've seen so far from drops out of a spore syringe. Should I let it grow before doing anything with it?


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Re: Question about multispores [Re: bolbol]
    #19508793 - 02/01/14 11:58 PM (9 years, 11 months ago)

Quote:

bolbol said:
what is so significant about rhizo growth of mycelium?
I had a search done and only results I got was that mycelium grows like that because of being in search of more nutrients and that its a bad sign??
my first set of jars inoculated with mexican cubes had normal growth and the new ones that i have with PES amazonian have the rhizo growth



You want to isolate rhizo growth so you can have the strongest and fastest growing strain if I'm not mistaken. I'm just now getting into isolation and cloning on agar but isn't that what people are after?


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Re: Question about multispores [Re: VYBE]
    #19508800 - 02/02/14 12:00 AM (9 years, 11 months ago)

yea that's what we are after. I wait for to pieces of mycelium to touch and they make rhizo? or that is how I begin isolation at least is what I understand.


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Re: Question about multispores [Re: Fert Nitty]
    #19508855 - 02/02/14 12:18 AM (9 years, 11 months ago)

You should look up an isolation and cloning tek. Not too difficult to understand but it's too much information for me to try to explain right now. Let me find a good link....

Here's sorta what we're talking about
https://mycotopia.net/topic/25864-strain-isolation-can-cubensis-benefit-from-advanced-teks/

There's also some links to some info by RR that I thought I had book marked but i can't find them


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Re: Question about multispores [Re: VYBE]
    #19534780 - 02/07/14 10:43 AM (9 years, 11 months ago)

I used liquid from a spore syringe on this agar plate so it kind of made a mess and created uneven mycelium growth all over the place. I was wondering if I took a hunk (in a s.a.b. of course) of myc. with tweezers and set it in the center of another agar plate, it should be more even growth.



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Re: Question about multispores [Re: Fert Nitty]
    #19534807 - 02/07/14 10:49 AM (9 years, 11 months ago)

Quote:

Fert Nitty said:
yea that's what we are after. I wait for to pieces of mycelium to touch and they make rhizo? or that is how I begin isolation at least is what I understand.




you're not necessarily after rhizo growth. On a petri dish rhizo growth is an indicator that you have good growth but it's not positive proof you have good genetics. Rhizo growth and tomentose growth are both dikaryotic mycelium IE the resultant of two monokaryons or hyphae or primary mycelium which is what a spore creates when it germinates. For practical purposes all the mycelium you see on your dish is dikaryotic mycelium which will usually show up tomentose on the first dish and then you may see rhizo growth on subsequent dishes. Rhizo growth is a good place to start when you're finding "Blind" isolates IE you don't know how they perform til you test them out. You could end up with shit or end up with gold and anything inbetween. If you're just starting out with agar don't even worry about isolates. Just put spores to agar transfer one or two times til you are sure you have clean growth and then use that, we call that sometimes limited MS or limited genetics. Then clone a good fruit from that grow. Worry about isolates when you've run out of fun with that.


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Re: Question about multispores [Re: bodhisatta]
    #19534833 - 02/07/14 10:57 AM (9 years, 11 months ago)

Im gonna transfer some of this to another plate.


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Re: Question about multispores [Re: Fert Nitty]
    #19534867 - 02/07/14 11:06 AM (9 years, 11 months ago)

:thumbup:


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Re: Question about multispores [Re: bodhisatta]
    #19535186 - 02/07/14 12:38 PM (9 years, 11 months ago)

will it work if u dont use agar at all and just pick a fruit, get the tissue at the base of the stem and just throw it in a grain jar?


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Re: Question about multispores [Re: bolbol]
    #19535322 - 02/07/14 01:21 PM (9 years, 11 months ago)

Seems to me throwing a chunk of mushroom in a grain jar to inoculate would have a high chance of contamination. 
Additionally, starting from just one little spot would probably mean slow colonization.


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Re: Question about multispores [Re: sourdoug]
    #19535533 - 02/07/14 02:29 PM (9 years, 11 months ago)

Ive inoculated a whole jar with a tiny cut from a healthy part of a cake that was half spread with contams and it went really good. Slow but it was okay.


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Re: Question about multispores [Re: bolbol]
    #19535580 - 02/07/14 02:42 PM (9 years, 11 months ago)

Quote:

bolbol said:
will it work if u dont use agar at all and just pick a fruit, get the tissue at the base of the stem and just throw it in a grain jar?



that would be instant fail. The mushroom is covered in contamination. You're throwing it into a sterile grain jar. EVEN ONE rogue contam will ruin your jar and the mushroom is covered in thousands. Even when cloning tissue to agar you take a small sliver from the middle of the fruit by ripping it in half. You don't cut the fruit because the blade will drag contams from the outside through the flesh. Even then you still might get contams on your first petri dish. Thats why we use the petri dishes to avoid failing hard in our grain jars.



Quote:

bolbol said:
Ive inoculated a whole jar with a tiny cut from a healthy part of a cake that was half spread with contams and it went really good. Slow but it was okay.



:smbfacepalm:


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Re: Question about multispores [Re: bodhisatta]
    #19535593 - 02/07/14 02:47 PM (9 years, 11 months ago)

Quote:

bodhisatta said:
Quote:

bolbol said:
will it work if u dont use agar at all and just pick a fruit, get the tissue at the base of the stem and just throw it in a grain jar?



that would be instant fail.



Not necessarily.....I did it once and succeeded.......but I used 10 tissue samples and knocked up 10 different grain jars.....only one made it without contamming.

Second link in my signature.


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Re: Question about multispores [Re: PussyFart]
    #19535609 - 02/07/14 02:51 PM (9 years, 11 months ago)

Quote:

PussyFart said:
Quote:

bodhisatta said:
Quote:

bolbol said:
will it work if u dont use agar at all and just pick a fruit, get the tissue at the base of the stem and just throw it in a grain jar?



that would be instant fail.



Not necessarily.....I did it once and succeeded.......but I used 10 tissue samples and knocked up 10 different grain jars.....only one made it without contamming.

Second link in my signature.



:lol:
For all intensive purpose I consider instant fail 10% or less success rates from now on.

Maybe not instant fail then I guess, but if you're going for a 10% success rate you can try it


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Re: Question about multispores [Re: bodhisatta]
    #19535616 - 02/07/14 02:52 PM (9 years, 11 months ago)

:lol:


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Re: Question about multispores [Re: bodhisatta]
    #19535702 - 02/07/14 03:16 PM (9 years, 11 months ago)

Yeah I meant the tissue inside of the stem and there shouldnt be any contams inside of it.

Quote:

Not necessarily.....I did it once and succeeded.......but I used 10 tissue samples and knocked up 10 different grain jars.....only one made it without contamming.

Second link in my signature.




Thats good enough for me to give it a try then :thumbup:


Edited by bolbol (02/07/14 03:22 PM)


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Re: Question about multispores [Re: bolbol]
    #19535774 - 02/07/14 03:29 PM (9 years, 11 months ago)

If you're going to try it, like bod said, tear the fruit, and be sure to flame sterilize whatever tool you choose to transfer the tissue, and do it in a SAB.


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Offlinebolbol
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Re: Question about multispores [Re: 1down5up]
    #19535787 - 02/07/14 03:32 PM (9 years, 11 months ago)

yeah I know. Thats what I was planning to do all along.:thumbup:


Edited by bolbol (02/07/14 03:33 PM)


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OfflineFert Nitty
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Re: Question about multispores [Re: bolbol]
    #19537021 - 02/07/14 09:24 PM (9 years, 11 months ago)

I just picked up a big bag of WBS and im gonna inoculate some via this agar plate and some vie MS. I read a good tek on here for spore prints where you start with one of the 1/2 pint jars sterile and transfer a cap into said jar. you can add water froma syringe through the inoc hole on top of the jar and even draw up the whole solution without opening the jar. Ive made so many syringes this way with almost zero contams. anything else im just gonna wait on my petri dishes.


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