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OfflineNightPuma1
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What is the point in boiling your agar first?
    #26652797 - 05/06/20 11:48 AM (3 years, 9 months ago)

Why do you sit there and mix together your agar and nutrients and boil them when they are about to get in the PC for 20 minutes (which is going to be a far higher temperature than boiling)?

Isn't whatever it is going to do going to do it in the bottle as well?


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InvisibleHtaeh
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Re: What is the point in boiling your agar first? [Re: NightPuma1]
    #26652806 - 05/06/20 11:51 AM (3 years, 9 months ago)

Mixing all the constituents together consistently to have a uniform liquid?


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Invisiblealaskappalachian
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Re: What is the point in boiling your agar first? [Re: NightPuma1]
    #26652817 - 05/06/20 11:56 AM (3 years, 9 months ago)

You're just getting the agar/nutes mixed/dissolved evenly.  You don't have to bring it to a boil.  I just mix everything cold and keep er stirring while I let the heat crank up, but I'm not looking to bring it to a boil.  Just heating and stirring until it's changed consistency/dissolved properly.


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OfflineOne of Us
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Re: What is the point in boiling your agar first? [Re: NightPuma1]
    #26652831 - 05/06/20 12:02 PM (3 years, 9 months ago)

As I see it,
People do this to make sure everything is dissolved properly and in solution before PCing. It gives the opportunity to stir or shake at a high temp to really help everything dissolve.

If all of the agar/MEA or whatever is in a clump at the bottom of the vessel it is more likely to caramelize, or otherwise become insoluble (during PC), so poured plates aren't as clear. At least, that was my thinking behind it.

I have, however, prepared agar without pre-boiling a few times recently and found little difference in clarity, so :shrug:


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OfflineNightPuma1
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Re: What is the point in boiling your agar first? [Re: One of Us]
    #26652837 - 05/06/20 12:08 PM (3 years, 9 months ago)

Okay I appreciate it guys but every time I do it I end up with a ton of sediment at the bottom (i.e. It is almost like there was no point in doing the boil at all).

What can I do to alleviate this?


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InvisibleHtaeh
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Re: What is the point in boiling your agar first? [Re: NightPuma1]
    #26652840 - 05/06/20 12:11 PM (3 years, 9 months ago)

What you using for nutrition?


--------------------
"To write is to wage war" ~ Voltaire
"The Drug Medical System cannot bear examination. To explain it would be to destroy it.." ~ R.T. Trall M.D.
"A single lie destroys a whole reputation for integrity." ~ Baltasar Gracian (1601-1658), Spanish philosopher 
"We'll know our disinformation program is complete when everything the US public believes is false."  ~  William J. Casey, 1981
"Fear always works to influence the population, when the population is uneducated."
"The secret of freedom lies in educating people, the secret of tyranny is in keeping them ignorant." ~  Maximilien Robespierre
"Knowledge makes a man unfit to be a slave."  ~  Frederick Douglass


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OfflineOne of Us
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Re: What is the point in boiling your agar first? [Re: Htaeh]
    #26652858 - 05/06/20 12:17 PM (3 years, 9 months ago)

You could always run the mix through a coffee filter just before PCing. That might help


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InvisibleHtaeh
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Re: What is the point in boiling your agar first? [Re: One of Us]
    #26652862 - 05/06/20 12:20 PM (3 years, 9 months ago)

That's true, I asked what nutrition because the same would happen to me using a potato I dried via dehydrator then blended for flakes. The sediment didn't make it into the plates mostly and the myc still grew as normal. I then switched to brf for nutrition and that seems to be much better imo.


--------------------
"To write is to wage war" ~ Voltaire
"The Drug Medical System cannot bear examination. To explain it would be to destroy it.." ~ R.T. Trall M.D.
"A single lie destroys a whole reputation for integrity." ~ Baltasar Gracian (1601-1658), Spanish philosopher 
"We'll know our disinformation program is complete when everything the US public believes is false."  ~  William J. Casey, 1981
"Fear always works to influence the population, when the population is uneducated."
"The secret of freedom lies in educating people, the secret of tyranny is in keeping them ignorant." ~  Maximilien Robespierre
"Knowledge makes a man unfit to be a slave."  ~  Frederick Douglass


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OfflineBrian Jones
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Re: What is the point in boiling your agar first? [Re: Htaeh]
    #26652871 - 05/06/20 12:25 PM (3 years, 9 months ago)

You will get some sediment with BRF too. If you use LME, and go through the nuking and shaking, it will be clear before it goes in the PC.


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OfflineAmino
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Re: What is the point in boiling your agar first? [Re: Htaeh]
    #26652876 - 05/06/20 12:28 PM (3 years, 9 months ago)

Agar takes ages to dissolve without stirring, malt extract is even worse, ensuring its all dissolved before hand leave a homogeneous solution. No lumps or clumps :smile:


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OfflineNightPuma1
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Re: What is the point in boiling your agar first? [Re: Amino]
    #26652891 - 05/06/20 12:42 PM (3 years, 9 months ago)

I have been going mad doing agar for days now. I think I've done 60 plates with no growth (although about 20 of those is too soon to tell).

The first recipe I was doing for a while did not result in any sediment (7.5 g Light Malt Extract, 10 g Agar Agar,
500 ml water). But it is also failed a lot so I think something is wrong (my LME is like 2 years old). I've used 3 different syringes from a very reputable source.

Yesterday I tried 25 grams Yellow Corn Meal, 3 grams dextrose, 10 grams agar, 500 mL water. It had a ton of sediment so I ended up pouring a lot fewer future dishes.

Today I'm trying 10 grams agar, 10 grams instant potatoes, 7 grams dextrose. Looks like a ton of sediment as well and I have nothing to filter it with ��

Procedure: still air box everything wiped down with tons of alcohol (again my problem is no growth not contamination). Put one droplet right in the center of the petri dish, then I hold the dish slightly diagonally to let the drop run a little bit (I don't feel comfortable doing an inoculation loop so this is at least a little bit of spreading it out.

Yes I shake the bejesus out of the syringes before use. I get nothing growing on any of these. I've used probably 4-5 syringes from a reputable source whose syringes have always worked on other mediums.

Perhaps my LME is too old which explains my failure in my first few runs - has anyone used more LME than just 7.5 grams in their agar recipe?

Thanks.


Edited by NightPuma1 (05/06/20 12:48 PM)


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Offlinehuangdeez
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Re: What is the point in boiling your agar first? [Re: NightPuma1]
    #26652946 - 05/06/20 01:21 PM (3 years, 9 months ago)

Bro, I had the same issue when I first tried agar. I made a shit load of plates (over 40) with 3 different agar recipes, and I got no growth. As soon as I used a print instead of syringes I got germination. I was also later able to get growth from a syringe by squirting the syringe on a sterile cotton swab, and then swabbing the plate. For some reason Just dropping spore solution to agar never worked for me either. Hope this helps. Also I never use that much agar for 500 ml I use 4tbsp agar 4tbsp nutes and 500 ML of water. Sometimes I add like a touch extra of agar, so 4-5 tbsp is plenty. Softer agar helps with germinating IME.


Edited by huangdeez (05/06/20 01:23 PM)


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OfflineA.k.aM
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Re: What is the point in boiling your agar first? [Re: huangdeez]
    #26652965 - 05/06/20 01:32 PM (3 years, 9 months ago)

I use lme and if I don’t heat it before pc it’ll have tons of sediment in there.

Microwaved and shake it first and its crystal clear.


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InvisibleSmartattack
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Re: What is the point in boiling your agar first? [Re: A.k.a]
    #26653044 - 05/06/20 02:14 PM (3 years, 9 months ago)

Sediment doesn't matter, but if you want clear plates just let it rest after mixing / heating, then decant and filter for crystal.


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