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OfflineDr. Sporesly
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Where to cut on agar for the best transfers?
    #26716188 - 06/03/20 09:01 AM (3 years, 7 months ago)

Hey all,

Finally got some decent agar growth and it's time to transfer it to continue the isolation process.

From the pictures below, what plates would you suggest cutting from, and at which location, size, etc.?

Thanks!



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InvisibleRoger Clemency
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Re: Where to cut on agar for the best transfers? [Re: Dr. Sporesly]
    #26716196 - 06/03/20 09:06 AM (3 years, 7 months ago)

Plates 2 and 4 at 7 o’clock. And plate 3 at 2:30

Just preference though. You could grab from just about anywhere on any of them except maybe 5. Looks off and a lil overgrown. All are getting to where you should transfer right now or a couple days ago. Lookin good though


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OfflineBallzagna
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Re: Where to cut on agar for the best transfers? [Re: Dr. Sporesly]
    #26716198 - 06/03/20 09:06 AM (3 years, 7 months ago)

1) probably in the 1-2 o'clock region

2) the thick bit at just above 3 o'clock

3) thick part at 2

4) thick part between 5 and 6

5) hard to say, kind of a messy plate so anywhere that looks clean

https://www.shroomery.org/forums/showflat.php/Number/18430998/fpart/all/vc/1

great guide for cleaning/isolating on agar


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OfflineDr. Sporesly
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Re: Where to cut on agar for the best transfers? [Re: Ballzagna]
    #26716207 - 06/03/20 09:11 AM (3 years, 7 months ago)

Thanks all!

Yeah I've read through the guide a few times but wanted to be accurate in what I do to have a higher success rate.

Out of 40 plates I poured, 39 were contaminated and only one showed growth (from syringe and loop)-

Took that plate and made these so really trying to hedge my bets haha


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InvisibleRoger Clemency
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Re: Where to cut on agar for the best transfers? [Re: Dr. Sporesly]
    #26716318 - 06/03/20 09:51 AM (3 years, 7 months ago)

Did you use a standard SAB? 39 of 40 almost seems like a sterilization failure and you got randomly lucky on one plate. Unless you were pouring them in open air or doing crazy things and rattling the SAB or something.

Or do you mean the myc didn’t germinate on bacteria or mold from a syringe? We’re there contaminants where you inoculated or was it satellite colonies in places you never touched?


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OfflineMarxcelium
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Re: Where to cut on agar for the best transfers? [Re: Ballzagna]
    #26716508 - 06/03/20 11:16 AM (3 years, 7 months ago)

Thanks for dropping this link! Been watching my first plates from spores grow and not sure how to attack a transfer, but I feel much more confident now.


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OfflineDr. Sporesly
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Re: Where to cut on agar for the best transfers? [Re: Roger Clemency]
    #26716934 - 06/03/20 02:09 PM (3 years, 7 months ago)

Quote:

Roger Clemency said:
Did you use a standard SAB? 39 of 40 almost seems like a sterilization failure and you got randomly lucky on one plate. Unless you were pouring them in open air or doing crazy things and rattling the SAB or something.

Or do you mean the myc didn’t germinate on bacteria or mold from a syringe? We’re there contaminants where you inoculated or was it satellite colonies in places you never touched?





Everything I've ever done has been an incredibly sterile environment inside an SAB and a completely sterilized room (can't get a flowhood right now).

I've poured ~80 plates and used a few different APE syringes from a reputable vendor. The first 40 were all bacteria (looks like cum on agar, yuck). Then the second bout was the one that worked.

I'd be surprised if it was a sterilization issue; however, that could be the case. But I've had no problems with the dozen+ other plates I've cloned from that one agar plate that was successful and I've never had any contamination otherwise – ever.

I think it's dirty syringes, but who knows.


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OfflineRogerRabbitM
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Re: Where to cut on agar for the best transfers? [Re: Dr. Sporesly]
    #26716999 - 06/03/20 02:55 PM (3 years, 7 months ago)

It's best to use a spore print, not syringe, when inoculating agar.  By swiping the spores in a zig-zag motion, you can see individual areas germinate and differentiate them from each other and contaminants before your first transfer(s).

I've seen spore syringes which would inoculate grains or BRF cakes successfully, but would contaminate with bacteria every time on agar.
RR


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InvisibleRoger Clemency
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Re: Where to cut on agar for the best transfers? [Re: RogerRabbit]
    #26717246 - 06/03/20 04:37 PM (3 years, 7 months ago)

If the nasty stuff only shows up where you inoculated then it's definitely the syringe. I hate syringes lol. I've always had a tough time trying to germinate on agar. The couple cake grows ive done with syringes went well though, like it's a magic combo.


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OfflineDr. Sporesly
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Registered: 05/20/20
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Re: Where to cut on agar for the best transfers? [Re: Roger Clemency]
    #26717472 - 06/03/20 06:46 PM (3 years, 7 months ago)

Quote:

RogerRabbit said:
It's best to use a spore print, not syringe, when inoculating agar.  By swiping the spores in a zig-zag motion, you can see individual areas germinate and differentiate them from each other and contaminants before your first transfer(s).

I've seen spore syringes which would inoculate grains or BRF cakes successfully, but would contaminate with bacteria every time on agar.
RR




Thanks for the response RR! Unfortunately there were no swabs available so I went the route of the syringe. I got lucky with the one plate so want to keep it going if I can.

Because the syringes are likely contaminated, other members suggested trying them on BRF cakes to see if they can work that way so I'm trying that now but it's going to take a while.


Quote:

Roger Clemency said:
If the nasty stuff only shows up where you inoculated then it's definitely the syringe. I hate syringes lol. I've always had a tough time trying to germinate on agar. The couple cake grows ive done with syringes went well though, like it's a magic combo.




^ Yeah I'm almost 100% sure it's the syringes.


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Offlineidontknow1
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Registered: 03/11/15
Posts: 39
Last seen: 2 months, 18 days
Re: Where to cut on agar for the best transfers? [Re: Dr. Sporesly]
    #26720019 - 06/04/20 07:08 PM (3 years, 7 months ago)

I've tried multiple APE syringes from a reputable vendor and never had success. Inoculating BRF cakes has never failed until I tried APE, germinated gray mold in 2 days over and over. I've heard APE is an unstable variety, perhaps this is the result of that.


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