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Catattack some Agar 3.0


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#261 Heirloom

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Posted 08 June 2017 - 12:02 AM

Thanks Cat for all the effort you put into this thread, lot of knowledge here. Glad I re read this before it got much longer.
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#262 CatsAndBats

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Posted 08 June 2017 - 01:27 PM

Thanks Cat for all the effort you put into this thread, lot of knowledge here. Glad I re read this before it got much longer.

 

 

Thanks man, I really appreciate it. :biggrin:



#263 Arathu

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Posted 09 June 2017 - 09:17 AM

Agar melting in the pot as I type.................it's aqua colored...........gotta go check the hood..............

 

A


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#264 Arathu

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Posted 09 June 2017 - 11:01 AM

And I WILL have a heated mag stirrer as soon as possible......it's just going to happen................... :cool:

 

I hate melting down previously set up agar in a double boiler........I always get chunks..............but I got 18 decent plates that are now cooling in the hood.......

 

Some transfers are going to happen in the witching hours of the full moon.............

 

A


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#265 CatsAndBats

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Posted 09 June 2017 - 11:04 AM

And I WILL have a heated mag stirrer as soon as possible......it's just going to happen................... :cool:

 

I hate melting down previously set up agar in a double boiler........I always get chunks..............but I got 18 decent plates that are now cooling in the hood.......

 

Some transfers are going to happen in the witching hours of the full moon.............

 

A

 

 

Why the stirrer? I made one for LC but didn't find any benefit.



#266 Arathu

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Posted 09 June 2017 - 11:07 AM

To get a nice smooth consistency to the agar when remelting it.........that or I was just rushing and should have melted it longer.............I never have problems like this with jelly jars..........hahahahahaha

 

It's just another skill set I'm working on........I got jelly jars down to an art form..........I want to get plates now........

 

It's just me bitching at myself in public............. :biggrin:

 

A


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#267 Microbe

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Posted 09 June 2017 - 12:03 PM

Arathu you are going to enjoy working with plates. I never have to remelt agar as i prepare only enough for what i need but a stir plate is recommended when remelting agar.

The greatest challenge with plates is learning to deal with condensation. Condensation is not a contamination vector as many claim. If there is a contamination, it will show itself with or without condensation dripping down on the agar surface. Its about being able to see and while stacking plates works it is time consuming and even after an hour or two i will still have some condensation on my plates.

Im will continue to stack to save space but im going to start using a heat gun. I have way to much shit to do everyday to wait around for hours to wait for plates to dry.

In processional labs they will actually slide theur lids slightly off. I have a nice flow hood but at the end of the day it is homemade and my lab is in my garage so i dont have the confidence to try that method.

Back to condensation. If you can get it drop back down to the surface, it will almost be absorbed back into the agar. However there are times when the moisture does not get absorbed and the problem here is that mycelium will grow much slower on wet surfaces then dry. If you were testing a culture this could give you a impression of sectoring, or non uniform growth. This is really no big deal as you will able to develop a trained eye to determine that.

The greatest concern about condensation that dripped down to the surface is that it can be a highway for bacteria to enter, leave, or move back and forth between colonies. Again its not a contamination vector, or the moisture doesnt result in the bacteria.

This is a problem when you are trying to clean up a dirty culture from bacterial infection. The bacteria can move across this moisture ahead of the colony and then be grown over before any signs of the bacteria are noticed thus transferring the bacteria yet again with what appears to be a healthy culture.

It is best practice prevent condensation. There is other shit like air bubbles, trapped in the middle of the agar and it will look like spots of mold underneath your colony when you hold it up to the light.

I have no idea how he finds time to do it but you have a good teacher. If you ever have any questions i too can help you out if you ever need it. I have a lot of experience in the lab myself and will try to explain the why behind the why.








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#268 Arathu

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Posted 09 June 2017 - 12:38 PM

I didn't want to remelt agar but the run cap on my blower shit itself in the middle of making agar......so I stashed the agar I was going to pour in the fridge.......

 

And since there was nothing visibly growing on it or in it I decided to melt it and pour.....

 

The next batch will go from the PC into plates........

 

Like I said I'm just being my own critic in public....it makes me better at what I decide to do..........

 

Yeah, I agree...my teacher knows his shit big time.......I'm fortunate.......

 

A


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#269 Microbe

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Posted 09 June 2017 - 02:13 PM

I didn't want to remelt agar but the run cap on my blower shit itself in the middle of making agar......so I stashed the agar I was going to pour in the fridge.......

And since there was nothing visibly growing on it or in it I decided to melt it and pour.....

The next batch will go from the PC into plates........

Like I said I'm just being my own critic in public....it makes me better at what I decide to do..........

Yeah, I agree...my teacher knows his shit big time.......I'm fortunate.......

A

Right on, i forgot your hood going down.

#270 Needles

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Posted 10 June 2017 - 10:35 AM

And I WILL have a heated mag stirrer as soon as possible......it's just going to happen................... :cool:
 
I hate melting down previously set up agar in a double boiler........I always get chunks..............but I got 18 decent plates that are now cooling in the hood.......
 
Some transfers are going to happen in the witching hours of the full moon.............
 
A


I find having a magnetic stirrer to be a valuable asset when working with agar. I have not tried a heated plate but have worked with hot liquids. There is a point where your liquid can take the amount of nutrients that you give it at cooler temperatures thus being saturated. If a liquid was heated it could absorb more of the nutrition and then being super saturated.
I find that a magnetic stirrer to be very useful when mixing dry ingredients. Just add a little at a time to prevent clumping then let it mix. I will sometimes let it mix for hours while I do other things. The most important reason for using a stirrer with agar is when you take agar containing vessels out of the PC. Using a gentle agitation with a slower speed, agar can be cooled and mixed at the same time. When agar has cooled off to the touch it can be poured in the plates without having to deal with air bubbles thus leaving you with a nice smooth surface that has equal amounts of your ingredients.
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#271 Arathu

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Posted 10 June 2017 - 11:13 AM

Well this batch isn't the finest and I won't be showing it off.............hahahahaha.......but I'm going to grow stuff on them anyway.........cause you gotta start somewhere.......... :biggrin:

 

And my jelly jars won't let me examine what's going on very well without opening them...........it's time for me to get professional......I have pitri's and I will order a shit ton more...........

 

Also my NEW AA-930 should be here Tuesday...........shit's getting real now......... :ohmy:

 

A


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#272 scott_1971_h

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Posted 13 June 2017 - 09:51 AM

Has anyone tried agar with antibiotics? Like, antibiotics included in the boil-up.

Maybe along with activated charcoal (theres no reason those 2 are mentioned together).

Could be super useful if you have a dodgy syringe or maybe your aseptic teks aren't your strongest point...

 



#273 CatsAndBats

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Posted 13 June 2017 - 10:56 AM

Has anyone tried agar with antibiotics? Like, antibiotics included in the boil-up.
Maybe along with activated charcoal (theres no reason those 2 are mentioned together).
Could be super useful if you have a dodgy syringe or maybe your aseptic teks aren't your strongest point...


I think the only antibiotic that can survive PCing is gentamicin:

en.m.wikipedia.org/wiki/Gentamicin

Fahtster has a salted agar tek too if I remember correctly.. it's on this thread somewhere methinks [I'm on my phone]

#274 scott_1971_h

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Posted 13 June 2017 - 11:31 AM

There's several:
https://groups.googl...kU/R2NmH4XZad8J
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