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OfflineSiLLySaNdMaN
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Registered: 08/12/19
Posts: 32
Last seen: 3 years, 8 months
2nd fail at Pastywhyte easy agar tek...
    #26335190 - 11/21/19 07:43 AM (4 years, 2 months ago)

I follow the tek to the T and each time they all get contaminated (twice). 😰. I understand that sterile technique is crucial. I flame sterilize needle, wipe down gloves and arms, pre crack lids, work swiftly, between each drop.

They all end up turning a peach/orange color

I don’t feel confident in trying a different agar tek. I like this one and everyone seems to have great success with it but wtf?

I’m following this tek: https://www.shroomery.org/forums/showflat.php/Number/23755440


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We are Gods, living in God’s world.


Edited by SiLLySaNdMaN (11/21/19 08:13 AM)


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OfflineStrohngBahd
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Registered: 04/15/19
Posts: 141
Loc: Desert
Last seen: 3 years, 9 months
Re: 2nd fail at Pastywhyte easy agar tek... [Re: SiLLySaNdMaN]
    #26335203 - 11/21/19 07:50 AM (4 years, 2 months ago)

Sounds like a bad syringe. I have had some that only shoot out bacteria. Contact who you got it from and see if it can be replaced. Unless you are in a place where it's legal to grow, I recommend saying something along the lines of "Looking at the spores through a microscope I noticed bacteria and/or mold".
Sounds like your sterile technique is good, you're not wiping the syringe with alcohol after flaming, right? 
Also just one drop per plate, and flame between drops/plates.


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No Turn Un-Stoned


Edited by StrohngBahd (11/21/19 07:51 AM)


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InvisibleTeaforTwo
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Registered: 09/22/19
Posts: 1,005
Loc: Psychonaut City, Mile Hi!
Re: 2nd fail at Pastywhyte easy agar tek... [Re: SiLLySaNdMaN]
    #26335211 - 11/21/19 07:54 AM (4 years, 2 months ago)

post some pics! 
you sure the color isn't from agar used?
you have your GE hole on top or side? one or two layer micropore tape?
folding paper towel and cover with foil too?
pc psi/time and how full loaded?
doing solid cone of steam vent for 10 minutes before shut valve/add weight and start session timer?

I haven't done pasties yet but from top of head think some of those responses may give better hints whats going on with yours:shrug:


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InvisibleSpunkyMonkey88
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Registered: 10/08/19
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Re: 2nd fail at Pastywhyte easy agar tek... [Re: SiLLySaNdMaN]
    #26335221 - 11/21/19 08:00 AM (4 years, 2 months ago)

Quote:

SiLLySaNdMaN said:
I follow the tek to the T and each time they all get contaminated (twice). 😰. I understand that sterile technique is crucial. I flame sterilize needle, wipe down gloves and arms, pre crack lids, work swiftly, between each drop.

They all end up turning a peach/orange color

I don’t feel confident in trying a different agar tek. I like this one and everyone seems to have great success with it but wtf?




Well its obviously got nothing to do with the agar... why do you feel you failed at the tek?

If you made a bunch of plates and dont open them and they contaminate then you failed the agar tek, your introducing contamination sometime after opening the plates, either by lousy technique or dirty innoculant...


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OfflineSiLLySaNdMaN
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Registered: 08/12/19
Posts: 32
Last seen: 3 years, 8 months
Re: 2nd fail at Pastywhyte easy agar tek... [Re: SpunkyMonkey88]
    #26335265 - 11/21/19 08:15 AM (4 years, 2 months ago)

Right right I know, not sure why I worded it like that. This is the tek I am following: https://www.shroomery.org/forums/showflat.php/Number/23755440


--------------------
We are Gods, living in God’s world.


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OfflineSiLLySaNdMaN
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Registered: 08/12/19
Posts: 32
Last seen: 3 years, 8 months
Re: 2nd fail at Pastywhyte easy agar tek... [Re: TeaforTwo]
    #26335286 - 11/21/19 08:23 AM (4 years, 2 months ago)

post some pics! 
you sure the color isn't from agar used? - well the agar is offwhite so I don’t think so
you have your GE hole on top or side? one or two layer micropore tape? On top plus 2 layers. You can get more than 1 cycle out of them, correct?
folding paper towel and cover with foil too? Correct
pc psi/time and how full loaded? Pc 45mins at 15psi of course. I’m doing 16 at a time to increase rate of success - not sure if this can be an issue. I’d hate to come off short
doing solid cone of steam vent for 10 minutes before shut valve/add weight and start session timer? Can you explain what this means? I leave the weight off until a valve pops up indicating it’s ready for the weight.

This is the tek I am following: https://www.shroomery.org/forums/showflat.php/Number/23755440


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We are Gods, living in God’s world.


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InvisibleTeaforTwo
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Registered: 09/22/19
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Re: 2nd fail at Pastywhyte easy agar tek... [Re: SiLLySaNdMaN]
    #26335408 - 11/21/19 09:12 AM (4 years, 2 months ago)

kk, some agar comes looking red, as Pastywhyte shows in his tek https://www.shroomery.org/forums/showflat.php/Number/19208976

yes, you can get many PC cycles out of them glad minis for no pours, as with other PP5 (container and lid must be pp5 tho). as long they dont deform or get really warped lids they are good.

and i just wanted to account for the peach/orange color you reported. so if not that, and not adding any food coloring and not the nutes added to agar making the color then I would fear bacterial contamination- but whole plate all plates now that's scary...

what type/brand/model of PC you using? not sure the valve popup thing, but basically you leave vent open until you have a solid cone of steam (for most types this is easy to see, mine doesnt use weights, its a control valve that vents downward due the deflector). once you have a nice cone of steam escaping set your timer for 10 minutes to ensure all air is vented out, replaced by superheated steam which is what we need to ensure sterility. after 10 minutes close valve or place weight on and set timer for your PC time recipe.

I see the tek you're following, looks like a good work flow incorporating pasties, we are still talking about your issue being on agar tho, correct?

and source of inoculate? not looking for vendor name if applicable, rather from spore print (vendor or friend), vendor syringe, swabs, clone slant, etc...

also, what variety shroom are you trying to grow?

as others mentioned this thread you may just have a bad syringe. another thought is do you ever leave a plate uninoculated as a control? as in just leave it alone fresh out the PC to see if anything grows (nothing should for weeks/months even if sterilized proper).


--------------------
See a mush cult pic you love? add it here: January 2020 PoM nominations thread! (up vote your faves to ensure they are included in final vote!). Community selected winner receives a 6mo Supporter account upgrade and/or other prizes from our awesome admin team!

Looking for something? Try Psilosopherr's Tek Compendium and pixelpopper's essentialzzz ForTheWin!

Be well, friend. Stay Strong and Beautiful, Always, and in All Ways~
Look Forward, Seek Truth, Find Real, Go Further~


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Offlinenicadim0
Stranger

Registered: 11/20/19
Posts: 1
Last seen: 4 years, 7 days
Re: 2nd fail at Pastywhyte easy agar tek... [Re: TeaforTwo]
    #26336496 - 11/21/19 05:26 PM (4 years, 2 months ago)

i read that pastywhyte's tek is outdated, and his PC times are too long


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Invisibleeatyualive
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Registered: 08/17/01
Posts: 19,026
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Re: 2nd fail at Pastywhyte easy agar tek... [Re: nicadim0]
    #26336928 - 11/21/19 08:30 PM (4 years, 2 months ago)

I only pc for 30 minutes at 15psi. Anyway do you have a spore print? You can make sterile swabs or buy them sterile and swab they print onto the agar. I prefer regular q tips. They are easy  to work with and I never use a spire loop anymore. If you try a print and it’s clean it might be the syringe.


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