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OfflineSpore211215
SporeGuy
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Registered: 06/06/16
Posts: 108
Last seen: 1 year, 8 months
Agar plates not getting clean transfers, need advise please
    #26567984 - 03/30/20 07:16 PM (3 years, 9 months ago)

OK boys and girls, I've been dabbling into agar and trying to get some clean plates to work off of but have been having some issues. Basically what it comes down to is i cant seem to transfer my plates without getting growth all over my new plate and i wanted to see if anyone here can tell me what I'm doing wrong.

I'll post as many relevant pics as i can to explain what I'm doing. The TEKs I've been using for my setup are as follows
Bod's Easy AF SAB TEK
Pastywhyte's Easy Agar Tek

The process I've been going through are as follows(My plates started as spore syringe to agar). So I start by spraying 70% ISO on my working surface and SAB then wiping down. I then bring my already grown agar plates into my SAB along with my PC'd and tin foiled clean agar plates. I proceed to put on my gloves, and spray my gloves and arms down with 70% ISO and inside my SAB to keep any particles in the air down. I then start to unwrap my clean plates to have a working stack ready to go and then once everything is unwrapped (but NOT opened/ having the lid off yet) i bring my arms out and light my propane torch. I then spray down my surgical knife and arms again before getting my blade red hot. Once i see its red i bring my arms back into the SAB and do a transfer like in this video(The agar transfer section). I then have my agar transfer complete on one new dish.

I like to try and do 2 transfers per dish so I place my lid back on the grown plate and get my blade and arms back outside the SAB. i then spray down the SAB with 70% ISO alog with my arms and blade and re heat my blade to make it clean again for more transfers. I repeat this process until I'm out of plates to work on. I have done about 3 transfers at this point and i keep getting messy plates and cant tell where I'm going wrong. Growth looks good to me but i just cant seem to get a good plate like i see everyone else getting.

Any advise? See where I'm screwing this up? If people want more pics or need any more info I'll be happy to give as much as I can. Big ups to all of you for helping me get this far and here to hoping I can proceed further, thanks!



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Invisiblestaytrippy420
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Registered: 03/23/13
Posts: 2,337
Loc: Canada Flag
Re: Agar plates not getting clean transfers, need advise please [Re: Spore211215]
    #26568104 - 03/30/20 08:36 PM (3 years, 9 months ago)

Ok it sounds like you got the basic procedure down. A few things I think are worth mentioning..

Using a wet hot towel under your sab and some sort of rack so your work isnt on the floor will help. The steam from the warm towel creates moisture on the walls and helps keep unwanted mold spores from bouncing around off the walls and floor. I always spray my sab down with isso before each session and everything that gets placed in the sab get wiped down as well.

Also are you double checking there’s no drafts in the room? Open windows, heats off, fans etc. You need to create still air by any means necessary. I brush my teeth before ever session and sometimes I go as far as wearing a dust mask so my breathing doesn’t stir shit up. Call me paranoid but when I started paying attention to these details and slowing down my movements in the sab my success rate went up.

i was also having trouble with the homemade no pour plates. I recommend you check these no mod no pour reusable containers out. They have better vision then the glads as well.
https://www.shroomery.org/forums/showflat.php/Number/26374264/page/1/fpart/1/vc/1


--------------------
Tek's I use

LAGM2020


Edited by staytrippy420 (03/30/20 08:44 PM)


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OfflineSpore211215
SporeGuy
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Registered: 06/06/16
Posts: 108
Last seen: 1 year, 8 months
Re: Agar plates not getting clean transfers, need advise please [Re: staytrippy420]
    #26568189 - 03/30/20 09:47 PM (3 years, 9 months ago)

OK, thanks for some helpful tips. Ill look into the hot towel since anything can help. And also them room does have a furnace vent that I was closing prior to working, but I'll just completely turn the heat down so there's no chance of any ventilation going on in the room. I might try a dust mask after all this corona stuff dies down to not screw up the chance of supplies to more important causes, but will definitely try it after. I'll attempt to slow down my movements like you said, and THANK you for the other TEK. the vision issues weren't a critical problem I was facing but it was just a major pain in the ass trying to see what was going on with my plates. Preciate all the help and I'll definitely send updates when the next round of transfers comes around (or if anything big comes up before then), Thanks!


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Offlinepoisoned
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Registered: 04/17/13
Posts: 1,738
Loc: Yurop
Last seen: 1 year, 3 months
Re: Agar plates not getting clean transfers, need advise please [Re: Spore211215]
    #26568411 - 03/31/20 03:48 AM (3 years, 9 months ago)

You don't need to iso everything each time you get your hands out of SAB. Especially not the insides of the SAB, that will basically introduce and push contaminants around instead of keeping the air inside still.

My procedure is as follows:
Wash my hands, set the SAB on the table (I just wipe the surface with a cloth before), spray the insides with soapy water, to get surfaces slightly misty. Don't wipe anything, those droplets will keep all the shit stuck to walls instead of falling onto your work. Then start putting items I need into the SAB, wiping everything with iso. Once done, turn on the torch, put the gloves on, wipe them with iso, burn the scalpel and go to work. I unwrap each plate individually as I go. The only reason why my hands are leaving the sub is to torch the blade. No touching of dirty iso bottle in the meantime.

Putting any significant amounts of iso into the SAB is actually relatively dangerous. The vapors can make you dizzy and can easily ignite or explode with that torch burning right next to it.

My contamination rate is ~5% and most of that is because I'm super sloppy sometimes. Also, I do my work next to a pile of dirty dishes quite regularly and they don't affect anything.


--------------------
How I do glass dishes


Edited by poisoned (03/31/20 03:51 AM)


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Invisiblestaytrippy420
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Registered: 03/23/13
Posts: 2,337
Loc: Canada Flag
Re: Agar plates not getting clean transfers, need advise please [Re: poisoned] * 1
    #26568631 - 03/31/20 08:39 AM (3 years, 9 months ago)

yeah I only spray the walls once and the steam from my hot towel does the rest.

Here is good sab conversation that’s already been had.
https://www.shroomery.org/forums/showflat.php/Number/26455740/fpart/1/vc/1


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OfflineSpore211215
SporeGuy
Male
Registered: 06/06/16
Posts: 108
Last seen: 1 year, 8 months
Re: Agar plates not getting clean transfers, need advise please [Re: staytrippy420]
    #26608709 - 04/17/20 05:59 PM (3 years, 9 months ago)

*UPDATE*

It turns out after letting my plates grow for an extended period of time that I was just growing gray mold that loved aerating its spores, no wonder I couldn't get a good transfer. Actually I was getting good transfers but I was just growing the wrong thing. :facepalm3:

I tossed everything and did a new set of like 24 THICC agar plates and inoculated about 20 with MS and 4 with a print that I transferred with a Q-tip. I went a little heavy handed on some of the MS plates so they aren't growing for shit, but everything else is having good growth. Hopefully I can post some good results in a few days but things are looking better this time around. Thanks for the advise and stay safe everyone!


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OfflineSpore211215
SporeGuy
Male
Registered: 06/06/16
Posts: 108
Last seen: 1 year, 8 months
Re: Agar plates not getting clean transfers, need advise please [Re: Spore211215]
    #26613691 - 04/19/20 07:33 PM (3 years, 9 months ago)

Pic update on second agar attempt. These are the best 7 plates out of the 24 I started with, I'll be doing transfers tomorrow and lets hope for the best.



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