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OfflineLuv_The_Cyans
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How to germ a tough swab?
    #26687613 - 05/22/20 08:41 AM (3 years, 8 months ago)

So I have been doing a lot of agar work the last few months. Very high success using various methods. I usually use an inoculation loop. I used a RW swab my first ever time with agar and all 6 dishes grew out strong. Last week I tried inoculating using sterile swabs from prints and all 25 plates show growth. My methods seems to be working over 100s of plates.
Now comes PE. I don't know if it is just me but my numbers seem to show otherwise. I have tried a PE-Uncut, PE-Melmac, 2 APE swabs with zero success. :frown: Seriously probably 50 plates and not a single one showed PE mycelium. About half did nothing and the other half eventually contaminated.
I tried lightly dragging the swab, roughly dragging, rolling, prodding, scraping off the tip into the dish, cutting off the whole tip and leaving it in the dish, putting tip in sterile water in a sterile shot glass and sucking up said solution into sterile syringe then placing on agar, low nute agar, soft agar, dog food agar, potatoe agar, grain water agar... and anything else I have seen on here. All work done in SAB with low contamination rate otherwise. Swabs came from 2 different vendors and both have sent replacements. I have more swabs and would like to get some PE going here. Can anyone please help me!


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Offlinetrip_hazard
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Re: How to germ a tough swab? [Re: Luv_The_Cyans] * 1
    #26687631 - 05/22/20 08:50 AM (3 years, 8 months ago)

This might be a stupid question, but if you've got PE's that you're swabbing why not just take a clone sample, clean it up, and put the myc to slant for long-term storage?


Edited by trip_hazard (05/22/20 08:50 AM)


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OfflineLuv_The_Cyans
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Re: How to germ a tough swab? [Re: trip_hazard]
    #26687693 - 05/22/20 09:15 AM (3 years, 8 months ago)

I don't have fruits. These are swabs from vendors. The only ones I have ever paid for and they don't work... :facepalm3:


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OfflineLuv_The_Cyans
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Re: How to germ a tough swab? [Re: Luv_The_Cyans]
    #26687923 - 05/22/20 11:22 AM (3 years, 8 months ago)

Hoping somebody has a tip to get these guys started? Maybe my samples are bad?? I don't like to blame my tools but I have had success with every other kind I have tried?


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Offlinetrip_hazard
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Re: How to germ a tough swab? [Re: Luv_The_Cyans] * 1
    #26689876 - 05/23/20 07:34 AM (3 years, 8 months ago)

PE's don't produce much in the way of spores so I'm a bit surprised vendors would be doing swabs.  Try contacting them and letting them know that the count was too low for microscopy and request a syringe instead - can't hurt to ask.


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InvisibleRiverBum
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Re: How to germ a tough swab? [Re: trip_hazard] * 1
    #26689943 - 05/23/20 08:18 AM (3 years, 8 months ago)

I have had the most success germinating pe swabs by prepping the agar soft then jamming the swab into the agar and putting a couple drops of sterile water on it. Works for me almost every time


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OnlineLogicaL ChaosM
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Re: How to germ a tough swab? [Re: RiverBum]
    #26689976 - 05/23/20 08:43 AM (3 years, 8 months ago)

Im writing this one down!

:takingnotes:


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OfflineMH5109
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Re: How to germ a tough swab? [Re: LogicaL Chaos]
    #26689994 - 05/23/20 08:53 AM (3 years, 8 months ago)

Iv got some rw spores I can't get to germ now im going to try a couple drops of water on the agar.


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Invisibleeatyualive
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Re: How to germ a tough swab? [Re: MH5109]
    #26690024 - 05/23/20 09:24 AM (3 years, 8 months ago)

Have you tried softer agar?


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OfflineLuv_The_Cyans
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Re: How to germ a tough swab? [Re: eatyualive]
    #26690053 - 05/23/20 09:49 AM (3 years, 8 months ago)

Yeah I tried 50% and 75% strength agar. Got slower growth with other swabs from a print but the PE did nothing. Do you have a suggested recipe to use? The vendors are from reddit, not shroomery sponsors. They have good reviews and stood behind their product. The prints I got from as a gift from same vendor did well.


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OfflineLuv_The_Cyans
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Re: How to germ a tough swab? [Re: Luv_The_Cyans]
    #26704249 - 05/29/20 10:48 AM (3 years, 7 months ago)

So finally some growth going from the APE! :bestcry:  The ones that didn't get covered in cobweb and did not do nothing had some nasty looking puffy slimy growth. It did not resemble cube mycelium but I saved 3 of them...just in case. 1 just turned into a nasty blob of contam but 2 of the others on the 75% strength agar eventually sectored off had an 1/4" area that looked more like cube mycelium. I transferred those 2 days ago and they seem to be growing out so far! :dancer:  Likely another transfer or 2 but I got APE  :poast:
Still no luck on the PEU but I am making PE progress.


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OfflineLuv_The_Cyans
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Re: How to germ a tough swab? [Re: Luv_The_Cyans]
    #26710898 - 06/01/20 08:30 AM (3 years, 7 months ago)

Tried out a Melmac swab I just got from a different source and no problems at all. I'm thinking I got bad swabs from 2 different sources :waitthatsbad: They backed their product and sent replacements but I was going a bit crazy there with all the failure. APE was a battle I am winning but its not over. PEU is still a losing cause...but it is nice to know it was likely not me! A bad tradesman blames his tools!


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