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Offlinegholladay
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Re: BOD's Comprehensive Agar TEK [Re: bodhisatta]
    #26198839 - 09/20/19 07:59 AM (4 years, 4 months ago)

Hey Bod! Thanks for all the great write ups. I poured my first grain water agar plates last night. got 12 out of 20. being a newbie i poured a little too much in each and ran out of agar. I have about 6 plastic petri dishes left over now. Is there anything those can be used for?

Also, what's your method for opening the sterile sleeve of petri dishes? do you open it outside of the SAB or inside? Your video shows nice stacks of ten.. i tried to open the plastic sleeve inside the SAB and it was a struggle.

Thanks!

GH


Edited by gholladay (09/20/19 08:02 AM)


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InvisiblebodhisattaMDiscordReddit
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Re: BOD's Comprehensive Agar TEK [Re: gholladay]
    #26198895 - 09/20/19 08:50 AM (4 years, 4 months ago)

I open it inside the box. It is a struggle till you get a method down. I hold it horizontal and slide them out then make the vertical ten stack


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Offlinegholladay
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Re: BOD's Comprehensive Agar TEK [Re: bodhisatta]
    #26205999 - 09/24/19 07:57 AM (4 years, 4 months ago)

Alright thank you. I'll try again. Despite the mess ups my agar plates seem to be clean so far. Nothing is growing on them after 5 days so i went ahead and dripped a drop from a new syringe on three of the dishes. Excited :smile:

GH


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Offlinemi.ro.ku
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Re: pour agar TEK [Re: CapitainFrog]
    #26216249 - 09/28/19 02:03 PM (4 years, 4 months ago)

thank you BOD!!


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OfflineLazerSounds
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Re: BOD's Comprehensive Agar TEK [Re: bodhisatta]
    #26218458 - 09/29/19 12:58 PM (4 years, 3 months ago)

I was wondering what the best temperature is to colonize your plates


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Re: BOD's Comprehensive Agar TEK [Re: LazerSounds]
    #26218486 - 09/29/19 01:17 PM (4 years, 3 months ago)

Room temperature


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OfflineLazerSounds
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Re: BOD's Comprehensive Agar TEK [Re: bodhisatta]
    #26220672 - 09/30/19 03:02 PM (4 years, 3 months ago)

Need help!
first time pouring plates they were too hot and they all were covered in condensation I tried the hot cup trick almost all of the condensation fell to the agar so I'm wondering and I know it's hard to tell from pictures but are these bacteria or water
Thanks for any help cheers


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OfflineLazerSounds
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Re: BOD's Comprehensive Agar TEK [Re: LazerSounds]
    #26220732 - 09/30/19 03:34 PM (4 years, 3 months ago)

More picks
I know that I fucked most of these up. I wasn't thinking and after flame sterilization with the wire tool I dipped it in isopropyl into the spore liquid onto the agar and tried the zig zag and drag tek The learning curve is tough 😂😂😂👊




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OfflineReptilianBrain
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Re: BOD's Comprehensive Agar TEK [Re: LazerSounds]
    #26221293 - 09/30/19 08:21 PM (4 years, 3 months ago)

My first attempt at agar (using the wrong type of agar: nutrient agar) is looking like this after 15 days. I have no idea what I’m looking at. It obviously doesn’t look like everyone else’s agar growth.


I’m still watching 5 plates of nutrient agar that all look pretty similar to the above pic, just in case they might be useful in some way. As of 4 days ago I started another 5 plates with PDAY agar, still no signs of anything with them.
Both batches have been kept at the same temp range of 75-80F. PC for 30-40 minutes, inoculated in SAB, wore gloves, mask, hairnet, plenty of 70%IPA wipedowns, lysoled air of the room, sprayed soap water inside sab, flamed syringe. Did everything as close to sterile tek as possible. Just curious, is this normal? Is it contamination? Any words of wisdom would be much appreciated.


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OfflineLazerSounds
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Re: BOD's Comprehensive Agar TEK [Re: ReptilianBrain]
    #26221354 - 09/30/19 09:12 PM (4 years, 3 months ago)

When you poured your plates was there condensation on the top after? Looks similar to some that I poured to hot?


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Invisiblestareatclouds
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Re: BOD's Comprehensive Agar TEK [Re: gholladay]
    #26221355 - 09/30/19 09:12 PM (4 years, 3 months ago)

Quote:

gholladay said:
Also, what's your method for opening the sterile sleeve of petri dishes? do you open it outside of the SAB or inside? Your video shows nice stacks of ten.. i tried to open the plastic sleeve inside the SAB and it was a struggle.





My SAB is just tall enough for the stack to sit barely (including blank plate on bottom to rise above towel). I use a flame sterilized blade to slice open the package and take out a 1-2 dishes so the stack is lower. Usually I just slowly scrunch up the sleeve until the bottom few are exposed while pinning the stack to the roof, then I slide those out and set them aside. Now the stack is low enough for me to scrunch the rest of the sleeve up comfortably. When another 5-7 are exposed, I stack those on the initial ones. And then there's more than enough room to get the rest out. I never pull open the sleeve. It's a hassle.

Anyway, that's the best way I've found to do it in a regular SAB. And then I made these for bags and pouring plates is 20x as easy. No condensation, no fight opening the stacks, can do more than 1 at a time, etc.



I wound up using the second version. I just put a few layers of tape where I wanted the arm holes for sturdiness and then cut them out. Going to write up a full Tek soon when I have a few more consistent bag runs with them as I'm still modifying it. But have done well over 20 bags to hit 100% and produce solid tubs with 2 or more flushes. I've seen people post for years about finding the biggest SABs or cutting and gluing 2 together or buying Plexiglass to make a custom one, etc. Not sure why nobody just flipped a big one on it's end and made the lid the face, but whatever. Could be a game changer, baby.


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InvisibleZero Nowhere
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Re: BOD's Comprehensive Agar TEK [Re: stareatclouds]
    #26221421 - 09/30/19 10:18 PM (4 years, 3 months ago)

Are the armholes on the side on the second one? One arm above the other?


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Re: BOD's Comprehensive Agar TEK [Re: Zero Nowhere]
    #26221445 - 09/30/19 10:39 PM (4 years, 3 months ago)

Sorry, that's kind of confusing. I didn't realize they were visible. That was my first SAB so the arm holes are drilled in the normal placements, but not in use for the modified version. They're completely taped over, so pretend they aren't even there. The arm holes are cut through the plastic cover where the lid should be like in the first picture. The box stands up on end and provides excellent height for bags and pouring. I'd prefer it wider, but whatever.

I'm quite sure I could just buy PVC pipe or something, make a tall rectangle with more width, and Saran wrap it all the same. Going to try that at some point.


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OfflineReptilianBrain
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Re: BOD's Comprehensive Agar TEK [Re: LazerSounds]
    #26221762 - 10/01/19 06:11 AM (4 years, 3 months ago)

I nocd about 15 minutes after the agar solidified, so I figured it was cool enough. But yeah, had a fair amount of condensation but it was mostly left over from trying to sterilize my plates. There was a bit more after pouring but started off with a bit already. My PDAY plates look great in terms of little/no condensation, but they haven’t started growing anything yet.


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Offlineghordly
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Re: BOD's Comprehensive Agar TEK [Re: ReptilianBrain] * 1
    #26224492 - 10/02/19 10:13 AM (4 years, 3 months ago)

Just want to take a moment to thank Bod for this amazing wealth of knowledge. Truly amazing work.


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I don't know what I'm doing but I think I've got somewhat of an idea..


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OfflineMrJeJo
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Re: pour agar TEK [Re: ghordly]
    #26229255 - 10/04/19 09:55 AM (4 years, 3 months ago)

Poured my first plates today. Created 400 ml agar, was a bit too much for the 20 60x15 plates I have. Used 400ml water with 8gr agar and 8gr LME. 25 min PC. Poured in SAB. Stored in ziplock bags. Some plates have condensation.



Edited by MrJeJo (10/04/19 09:56 AM)


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OfflineiEnter
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Re: pour agar TEK [Re: MrJeJo]
    #26238025 - 10/08/19 02:03 PM (4 years, 3 months ago)

Thanks for the awesome guide!

Just got my first plates colonized and I'm ready to do a transfer tomorrow.

What do you all think? Any issues?



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Offlinesoberami
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Re: BOD's Comprehensive Agar TEK [Re: bodhisatta]
    #26278604 - 10/26/19 03:05 PM (4 years, 3 months ago)

Thank you for this. I am still a little confused about the Malt and/or (or vs.?) brf. Which is the preferred, or do I use both (without the broth of grain soak water)?


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Offlinejbgtaa
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Re: pour agar TEK [Re: iEnter]
    #26278685 - 10/26/19 03:33 PM (4 years, 3 months ago)

Quote:

iEnter said:
Thanks for the awesome guide!

Just got my first plates colonized and I'm ready to do a transfer tomorrow.

What do you all think? Any issues?





This is bods thread so sorry bod but...
All those plates look like really solid germination. If you’re going to go to grain quickly, I’d take a transfer from each of the plates at a thick area of growth. If your trying to further isolate go for the leading edge of a very thin selection but make sure it looks really good. Good job, I’d just take 1 more transfer (2 if bacteria or mold pops up, but that’s a totally different problem)from each and put it to grain.


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InvisiblebodhisattaMDiscordReddit
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Re: pour agar TEK [Re: jbgtaa]
    #26279256 - 10/26/19 09:10 PM (4 years, 3 months ago)

Quote:

soberami said:
Thank you for this. I am still a little confused about the Malt and/or (or vs.?) brf. Which is the preferred, or do I use both (without the broth of grain soak water)?



Either

Quote:

jbgtaa said:
Quote:

iEnter said:
Thanks for the awesome guide!

Just got my first plates colonized and I'm ready to do a transfer tomorrow.

What do you all think? Any issues?





This is bods thread so sorry bod but...
All those plates look like really solid germination. If you’re going to go to grain quickly, I’d take a transfer from each of the plates at a thick area of growth. If your trying to further isolate go for the leading edge of a very thin selection but make sure it looks really good. Good job, I’d just take 1 more transfer (2 if bacteria or mold pops up, but that’s a totally different problem)from each and put it to grain.



No worries missed that post


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