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OfflineUmbraSprite
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PE spore syringe to agar....can anyone confirm?
    #26587186 - 04/08/20 07:52 PM (3 years, 10 months ago)

Recently picked up a PE spore syringe and inoculated two agar plates and six BRF 1/2 pint jars.

The growth was much faster than my experience with B  (visible in the jars by day 3) and a bit "fluffier" looking than what I am used to.  Further I got no growth on one plate (never had this happen before) and the second plate I checked today and a silver dollar sized growth resembling a ball of cotton had grown all the way up against the plate lid.  I am assuming this is the PE mycelium but given how different it looks from B  I am not ruling out that I somehow ended up with contaminants.  It's highly unlikely that I contaminated 6 jars and two plates given my background (microbiologist) and track record working with B  on agar.  That said...what I am seeing isn't what I expected.

Does anyone have any experience culturing PE on agar?  Could this silver dollar sized cottony mass be the PE?  Does PE mycelium look that morphologically different from B+?

Usually I have photo's but this time I trashed the plates before getting any. I did sample the "cottony mass" to a new plate so I should be able to get a shot of that in a few days.

Thanks in advance.


Edited by UmbraSprite (04/08/20 07:53 PM)


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Offlinerumfor69
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Re: PE spore syringe to agar....can anyone confirm? [Re: UmbraSprite]
    #26587285 - 04/08/20 08:34 PM (3 years, 10 months ago)

3 days does seem suspiciously fast for spore germination.

But yeah without pics it's difficult to give much opinion


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Offlineshevanel
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Re: PE spore syringe to agar....can anyone confirm? [Re: rumfor69]
    #26587314 - 04/08/20 08:45 PM (3 years, 10 months ago)

Ive seen plates [recently] germinate like clockwork on the 3rd if not 4th day so its not too terribly unusual.


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Offlinerumfor69
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Re: PE spore syringe to agar....can anyone confirm? [Re: shevanel]
    #26587323 - 04/08/20 08:48 PM (3 years, 10 months ago)

I was referring to the jars he said it was visible in 3 days


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OfflineUmbraSprite
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Re: PE spore syringe to agar....can anyone confirm? [Re: rumfor69]
    #26587336 - 04/08/20 08:51 PM (3 years, 10 months ago)

Grabbed the plate from the trash.  Looks like total mold to me but if so the syringe must have been contaminated.  I could write off a contaminated plate or two but for the jars to have (what looks like) similar growth would indicate this was not a technique issues.



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Offlinerumfor69
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Re: PE spore syringe to agar....can anyone confirm? [Re: UmbraSprite]
    #26587354 - 04/08/20 08:57 PM (3 years, 10 months ago)

Hmmm puffy, thick, fuzzy, fast growing...probably mold.

But the new plate you transferred to will tell more story.


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OfflineUmbraSprite
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Re: PE spore syringe to agar....can anyone confirm? [Re: rumfor69]
    #26587422 - 04/08/20 09:26 PM (3 years, 10 months ago)

How common is it to see contaminated syringes?


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Offlinerumfor69
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Re: PE spore syringe to agar....can anyone confirm? [Re: UmbraSprite]
    #26587524 - 04/08/20 10:06 PM (3 years, 10 months ago)

All spores are contaminated to some extent it's just in their nature.
They're grown in the open air so they get contams on them.


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OfflineUmbraSprite
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Re: PE spore syringe to agar....can anyone confirm? [Re: rumfor69]
    #26588339 - 04/09/20 08:54 AM (3 years, 10 months ago)

Well that sucks.  They are expensive and PE's are hard to come by in the first place.  Given that both plates and six different jars have this stuff and I sterilized the ports and needle between jars/plates I am highly inclined to believe this was a bad batch of spores.

Something else to note is that my agar was a uniform opaque last time but I suppose I ordered from a different vendor this time and there is a lot of heterogeneity in the agar.  Even a few solids.  At first I thought it was contaminated but if I am not mistaken depending on the way it is made (i.e. not from a powdered form) this can be the case and not have anything to do with contamination.  Either way it wouldn't explain the jars and the growth on the plates is isolated to where I inoculated.


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Offlinerumfor69
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Re: PE spore syringe to agar....can anyone confirm? [Re: UmbraSprite]
    #26588373 - 04/09/20 09:06 AM (3 years, 10 months ago)

I just get the cheap PDA off Amazon for $16 it seems to always work well.
I use 1/2 pint wide mouth jars with sfd's, holes in metal lid, and ring
setup. Maybe it was an  extra dirty syringe, that can happen too. Definitely
get syringes from a Shroomery sponsor through the community menu if
you didn't.

Wait on the newly transferred plate and if it doesn't work then get
another syringe or a print from the market forum if available. Make
up a dozen plates and put two drops on each one spaced apart to get
lots of dice roll chances to get something clean to transfer.


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OfflineB21
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Re: PE spore syringe to agar....can anyone confirm? [Re: UmbraSprite]
    #26588393 - 04/09/20 09:17 AM (3 years, 10 months ago)

Hello my friend..
Quote:

UmbraSprite said:
Well that sucks.  They are expensive and PE's are hard to come by in the first place.  Given that both plates and six different jars have this stuff and I sterilized the ports and needle between jars/plates I am highly inclined to believe this was a bad batch of spores.



Don’t think like that bro,,, Agar is a baaaaddd mfr. You should keep trying I think. The only thing that I could think of is where you got the syringe (don’t say any names) I just think that there are some people out there that don’t practice very sterile techniques when making syringes to sell. Like when I go through an online trusted vendor I very rarely see contams but when I bought a syringe off some dude on another site or app it would only yield yellow slime,,, like they just sucked toilet water into the syringe. Imo I’d birth one of your cakes (when it’s fully colonized) in a sab and put a tiny chunk of the cake on an agar plate,, this is only my opinion,,, like it’s what I would do. I’ve had trouble with syringes before and done what I just described and eventually had clean mycelium that went on to put out many beautiful fruits.
Don’t give up, keep trying, good luck, and most importantly HAVE FUN


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Edited by B21 (04/09/20 09:18 AM)


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OfflineB21
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Re: PE spore syringe to agar....can anyone confirm? [Re: UmbraSprite]
    #26588407 - 04/09/20 09:21 AM (3 years, 10 months ago)

Quote:

UmbraSprite said:

Does anyone have any experience culturing PE on agar?




https://www.shroomery.org/forums/showflat.php/Number/25022155/fpart/all/vc/1
Just something to check out. It’s really cool


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Edited by B21 (04/09/20 09:22 AM)


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OfflineUmbraSprite
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Re: PE spore syringe to agar....can anyone confirm? [Re: rumfor69]
    #26588432 - 04/09/20 09:33 AM (3 years, 10 months ago)

@rumfor69

I try and use the supporter vendors but would have to go back and look where I ended up.  I am pretty sure none of the vendors here had the spores I was looking for. 

I didn't use the entire syringe so I have suspension to try again but as I have been saying I am pretty confident this was a bad syringe of suspension.


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OfflineUmbraSprite
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Re: PE spore syringe to agar....can anyone confirm? [Re: UmbraSprite]
    #26588436 - 04/09/20 09:35 AM (3 years, 10 months ago)

@B21

Yeah the plates in that link look like what I am used to when culturing B+.  I didn't see why the mycelium would have such big morphological differences thus why I assumed contamination.  Further the growth I have grew vertically very far and very quickly.  The cubensis stays relatively low to the agar.

Oh and the vendor was a long standing supporter of this site. 


Edited by UmbraSprite (04/09/20 09:38 AM)


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OfflineA.k.aM
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Re: PE spore syringe to agar....can anyone confirm? [Re: UmbraSprite]
    #26588455 - 04/09/20 09:47 AM (3 years, 10 months ago)

Most spores I’ve used germ on day 4 seems like, but I’ve had some go off in one day.


PE myc has always been very fast and aggressive for me.

That said idk what’s on that plate. Could be super tomentose myc I guess.


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Offlinetrip_hazard
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Re: PE spore syringe to agar....can anyone confirm? [Re: UmbraSprite]
    #26588533 - 04/09/20 10:26 AM (3 years, 10 months ago)

Quote:

UmbraSprite said:
Does anyone have any experience culturing PE on agar? 




Just recently did some PE to agar from an older syringe; it took 7 days before there was any sign of germination.


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OfflineUmbraSprite
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Re: PE spore syringe to agar....can anyone confirm? [Re: trip_hazard]
    #26589730 - 04/09/20 09:07 PM (3 years, 10 months ago)

Mold IMO. Very robust, healthy mold loving all the nutrients.




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OfflineUmbraSprite
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Re: PE spore syringe to agar....can anyone confirm? [Re: UmbraSprite]
    #26590756 - 04/10/20 10:33 AM (3 years, 10 months ago)

Update: The vendor confirmed they had a bad batch of spores and has volunteered to replace them.


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OfflineUmbraSprite
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Re: PE spore syringe to agar....can anyone confirm? [Re: UmbraSprite]
    #26636493 - 04/29/20 10:29 AM (3 years, 9 months ago)

Hey all...

New plates are about 7 days old and I wanted to get some opinions.

I plated the suspect PE syringe, a replacement sent from the vendor, a syringe of PE spores from a new vendor and a plate transfer from my B+ clone. 

To be honest they all look about the same.  I haven't seen the huge, fast, puffy growth that tried to push the lid off of the plate however there is definitely some vertical growth.  Keep in mind I am pretty new at putting cubes on agar.  I have done tons of bacterial work over the years but not fungal.  It is 100% possible that what is growing is completely normal cube growth.

I will try and get some new photo's up today.  Hopefully a few of you are still following!


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OfflineUmbraSprite
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Re: PE spore syringe to agar....can anyone confirm? [Re: UmbraSprite]
    #26636517 - 04/29/20 10:43 AM (3 years, 9 months ago)

Alright.  8 days growth.

B+ plate transfer (original started from fruit tissue)



PE Spore Syringe #1 (new vendor)



PE Spore Syringe #2 (original vendor)



PE Spore Syringe #3 (original vendor)



Edited by UmbraSprite (04/29/20 10:47 AM)


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