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Offlineverum subsequentis
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Re: Bod's Comprehensive Agar TEK [Re: verum subsequentis] * 2
    #24225946 - 04/07/17 07:04 PM (3 years, 5 months ago)

OK, this is my first attempt at uploading pictures so...









First two are taken from the underside with a light behind but I couldn't really see it. So I sacrificed the third plate for the photo. The lighting really catches it and makes it seem really bad but without that perfect angle it is hardly noticeable.  Fourth is my led computer fan magnetic stirrer just for fun.


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InvisiblebodhisattaM
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Re: Bod's Comprehensive Agar TEK [Re: verum subsequentis]
    #24225992 - 04/07/17 07:26 PM (3 years, 5 months ago)

Looks like it could be bacteria to me. You don't happen to have a scope? I've never seen water dry up on a dish so hard water doesn't make sense to me. Petri dishes are 98% water so there's no evaporation leaving hard water residue behind


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Offlineverum subsequentis
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Re: Bod's Comprehensive Agar TEK [Re: bodhisatta]
    #24225998 - 04/07/17 07:30 PM (3 years, 5 months ago)

I don't have a scope but I'll be getting one. Did you just see it in the third picture or all Petris? It looks awfully bad in that picture for sure. I'm just not totally convinced it's bacteria but, I am admittedly not very familiar with it.


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InvisiblebodhisattaM
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Re: Bod's Comprehensive Agar TEK [Re: verum subsequentis]
    #24226007 - 04/07/17 07:33 PM (3 years, 5 months ago)

Third picture was the one I really saw it on


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Re: Bod's Comprehensive Agar TEK [Re: bodhisatta] * 1
    #24234917 - 04/11/17 03:29 PM (3 years, 5 months ago)



--------------------
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OfflineRoyalmensch
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Re: pour agar TEK [Re: bodhisatta]
    #24235927 - 04/11/17 10:47 PM (3 years, 5 months ago)

Noted, I'll have to revisit this when I'm ready.


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Re: pour agar TEK [Re: Royalmensch]
    #24271016 - 04/25/17 05:45 PM (3 years, 4 months ago)

bump


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Re: pour agar TEK [Re: bodhisatta]
    #24273593 - 04/26/17 05:21 PM (3 years, 4 months ago)

91 can be diluted to 70%
C1V1=C2V2 where c is concentration and v is volume
1L of 91% turns into 1.3L of 70% add 300mL of water to 1L of 91%


Quote:

bodhisatta said:
For iso and ethanol

70% kills better
It has the right osmotic pressure and tonicity to penetrate cells and denature their insides. It may denature surface proteins on cell walls and bacterial envelopes but not to the extent higher percentage alcohol(both iso and ethanol) will. Alcohol is able to permeate cells and then go to work on intracellular components

A secondary benefit is it evaporates slower which potentiates it's killing ability

91% doesn't have the right osmolality. It doesn't cross the cell membrane or bacterial envelope as well. It "puckers up" cells. It denatures the proteins on cell surfaces. This disables cells but doesn't necessarily kill them, they can re-able themselves later when conditions are favorable

A secondary hindrance of 91% is it evaporates faster making it even less effective than it already is.


Also you shouldn't be using it at incredible rates
A bottle should last a long time. If you go through it like water your sterile technique sucks ass and you're relying on chemicals too much. Chemicals won't save your ass




Quote:

RogerRabbit said:
70% is preferred, but it has nothing to do with rate of evaporation. 

Cells admit water through their cell walls via osmosis.  Cell walls are particularly good at preventing the entry of toxins, so by mixing water with the alcohol, it 'tricks' the cell wall into admitting the mixture, which then kills the cell as the alcohol evaporates back out.

I'm sure one of our resident chemists can put it in more scientific terms, but that's the jest of it.
RR




Quote:

RogerRabbit said:
It has nothing to do with how fast the alcohol evaporates. 

It has to do with pure alcohol not being admitted inside the cell walls by osmosis.  If water is mixed with the alcohol, it tricks the cell wall into admitting it, therefore killing the cell. 70% has been shown to be a good mixture for sanitizing, and superior to higher concentrations.
RR




Quote:

bodhisatta said:
Alcohols
Although several alcohols have been shown to be effective antimicrobials, ethyl alcohol (ethanol, alcohol), isopropyl alcohol (isopropanol, propan-2-ol) and n -propanol (in particular in Europe) are the most widely used (337). Alcohols exhibit rapid broad-spectrum antimicrobial activity against vegetative bacteria (including mycobacteria), viruses, and fungi but are not sporicidal. They are, however, known to inhibit sporulation and spore germination (545), but this effect is reversible (513). Because of the lack of sporicidal activity, alcohols are not recommended for sterilization but are widely used for both hard-surface disinfection and skin antisepsis. Lower concentrations may also be used as preservatives and to potentiate the activity of other biocides. Many alcohol products include low levels of other biocides (in particular chlorhexidine), which remain on the skin following evaporation of the alcohol, or excipients (including emollients), which decrease the evaporation time of the alcohol and can significantly increase product efficacy (68). In general, isopropyl alcohol is considered slightly more efficacious against bacteria (95) and ethyl alcohol is more potent against viruses (259); however, this is dependent on the concentrations of both the active agent and the test microorganism. For example, isopropyl alcohol has greater lipophilic properties than ethyl alcohol and is less active against hydrophilic viruses (e.g., poliovirus) (259). Generally, the antimicrobial activity of alcohols is significantly lower at concentrations below 50% and is optimal in the 60 to 90% range.

iso is preferred to ethanol as a sanitizer because iso solubilizes fats better than ethanol does, hence being slightly more bactericidal

at 70% concentration it's hypotonic look up tonicity and osmolality to understand why this is important

Anyway ethanol and iso are interchangeable as sanitizer against non viral contaminants

You can go on if you would like I'm done here with this. You can be into facts or not.
90+ will inhibit the cell wall and not penetrate the interior as well thus not being as effective.

70% has a more effective osmotic pressure than 71-90

Evaporation is a secondary benefit

91+ % iso and ethanol are inferior sanitizer to 70 iso and ethanol

there's a reason hospitals and labs use 70% and it's not cost.

first and foremost the osmotic pressure of the 70% concentration is best the tonicity of this concentration means it will be able to denature the inside of a cell rather than surface proteins on the bacterial envelope. If a bacterial envelop is disabled the cell can come back to life later on since the inside of the cell hasn't been scrambled by the alcohol.
the added benefit is that 70% doesn't evaporate as fast this potentates it's effectiveness but is not the cause for the effectiveness. 




--------------------
police violence is no accident
ACAB
There's two kinds of people. Those who have had a bad interaction with cops and those who haven't got their turn


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Offlineyojay50
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Re: pour agar TEK [Re: bodhisatta]
    #24290618 - 05/02/17 09:01 PM (3 years, 4 months ago)

Quote:

bodhisatta said:
91 can be diluted to 70%
C1V1=C2V2 where c is concentration and v is volume
1L of 91% turns into 1.3L of 70% add 300mL of water to 1L of 91%


Quote:

bodhisatta said:
For iso and ethanol

70% kills better
It has the right osmotic pressure and tonicity to penetrate cells and denature their insides. It may denature surface proteins on cell walls and bacterial envelopes but not to the extent higher percentage alcohol(both iso and ethanol) will. Alcohol is able to permeate cells and then go to work on intracellular components

A secondary benefit is it evaporates slower which potentiates it's killing ability

91% doesn't have the right osmolality. It doesn't cross the cell membrane or bacterial envelope as well. It "puckers up" cells. It denatures the proteins on cell surfaces. This disables cells but doesn't necessarily kill them, they can re-able themselves later when conditions are favorable

A secondary hindrance of 91% is it evaporates faster making it even less effective than it already is.


Also you shouldn't be using it at incredible rates
A bottle should last a long time. If you go through it like water your sterile technique sucks ass and you're relying on chemicals too much. Chemicals won't save your ass




Quote:

RogerRabbit said:
70% is preferred, but it has nothing to do with rate of evaporation. 

Cells admit water through their cell walls via osmosis.  Cell walls are particularly good at preventing the entry of toxins, so by mixing water with the alcohol, it 'tricks' the cell wall into admitting the mixture, which then kills the cell as the alcohol evaporates back out.

I'm sure one of our resident chemists can put it in more scientific terms, but that's the jest of it.
RR




Quote:

RogerRabbit said:
It has nothing to do with how fast the alcohol evaporates. 

It has to do with pure alcohol not being admitted inside the cell walls by osmosis.  If water is mixed with the alcohol, it tricks the cell wall into admitting it, therefore killing the cell. 70% has been shown to be a good mixture for sanitizing, and superior to higher concentrations.
RR




Quote:

bodhisatta said:
Alcohols
Although several alcohols have been shown to be effective antimicrobials, ethyl alcohol (ethanol, alcohol), isopropyl alcohol (isopropanol, propan-2-ol) and n -propanol (in particular in Europe) are the most widely used (337). Alcohols exhibit rapid broad-spectrum antimicrobial activity against vegetative bacteria (including mycobacteria), viruses, and fungi but are not sporicidal. They are, however, known to inhibit sporulation and spore germination (545), but this effect is reversible (513). Because of the lack of sporicidal activity, alcohols are not recommended for sterilization but are widely used for both hard-surface disinfection and skin antisepsis. Lower concentrations may also be used as preservatives and to potentiate the activity of other biocides. Many alcohol products include low levels of other biocides (in particular chlorhexidine), which remain on the skin following evaporation of the alcohol, or excipients (including emollients), which decrease the evaporation time of the alcohol and can significantly increase product efficacy (68). In general, isopropyl alcohol is considered slightly more efficacious against bacteria (95) and ethyl alcohol is more potent against viruses (259); however, this is dependent on the concentrations of both the active agent and the test microorganism. For example, isopropyl alcohol has greater lipophilic properties than ethyl alcohol and is less active against hydrophilic viruses (e.g., poliovirus) (259). Generally, the antimicrobial activity of alcohols is significantly lower at concentrations below 50% and is optimal in the 60 to 90% range.

iso is preferred to ethanol as a sanitizer because iso solubilizes fats better than ethanol does, hence being slightly more bactericidal

at 70% concentration it's hypotonic look up tonicity and osmolality to understand why this is important

Anyway ethanol and iso are interchangeable as sanitizer against non viral contaminants

You can go on if you would like I'm done here with this. You can be into facts or not.
90+ will inhibit the cell wall and not penetrate the interior as well thus not being as effective.

70% has a more effective osmotic pressure than 71-90

Evaporation is a secondary benefit

91+ % iso and ethanol are inferior sanitizer to 70 iso and ethanol

there's a reason hospitals and labs use 70% and it's not cost.

first and foremost the osmotic pressure of the 70% concentration is best the tonicity of this concentration means it will be able to denature the inside of a cell rather than surface proteins on the bacterial envelope. If a bacterial envelop is disabled the cell can come back to life later on since the inside of the cell hasn't been scrambled by the alcohol.
the added benefit is that 70% doesn't evaporate as fast this potentates it's effectiveness but is not the cause for the effectiveness. 







BEAUTIFUL  :takingnotes:

i used 91% to sterilize my syringe and more, might have a contam on my agar.


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Offlinegillaputty
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Re: pour agar TEK [Re: yojay50]
    #24330260 - 05/18/17 04:37 PM (3 years, 4 months ago)

Agar is sensitive around rye.  The prime example of contamination from other forms of germs that are on the unshucked rye.


Try using one of the better spore prints you might find for better rhizo.


Edited by gillaputty (05/18/17 04:37 PM)


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InvisiblebodhisattaM
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Re: pour agar TEK [Re: gillaputty]
    #24330312 - 05/18/17 04:50 PM (3 years, 4 months ago)

What?


--------------------
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ACAB
There's two kinds of people. Those who have had a bad interaction with cops and those who haven't got their turn


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Offlinemrmazdarx9
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Re: pour agar TEK [Re: yojay50]
    #24330329 - 05/18/17 04:55 PM (3 years, 4 months ago)

Quote:

yojay50 said:
Quote:

bodhisatta said:
91 can be diluted to 70%
C1V1=C2V2 where c is concentration and v is volume
1L of 91% turns into 1.3L of 70% add 300mL of water to 1L of 91%


Quote:

bodhisatta said:
For iso and ethanol

70% kills better
It has the right osmotic pressure and tonicity to penetrate cells and denature their insides. It may denature surface proteins on cell walls and bacterial envelopes but not to the extent higher percentage alcohol(both iso and ethanol) will. Alcohol is able to permeate cells and then go to work on intracellular components

A secondary benefit is it evaporates slower which potentiates it's killing ability

91% doesn't have the right osmolality. It doesn't cross the cell membrane or bacterial envelope as well. It "puckers up" cells. It denatures the proteins on cell surfaces. This disables cells but doesn't necessarily kill them, they can re-able themselves later when conditions are favorable

A secondary hindrance of 91% is it evaporates faster making it even less effective than it already is.


Also you shouldn't be using it at incredible rates
A bottle should last a long time. If you go through it like water your sterile technique sucks ass and you're relying on chemicals too much. Chemicals won't save your ass




Quote:

RogerRabbit said:
70% is preferred, but it has nothing to do with rate of evaporation. 

Cells admit water through their cell walls via osmosis.  Cell walls are particularly good at preventing the entry of toxins, so by mixing water with the alcohol, it 'tricks' the cell wall into admitting the mixture, which then kills the cell as the alcohol evaporates back out.

I'm sure one of our resident chemists can put it in more scientific terms, but that's the jest of it.
RR




Quote:

RogerRabbit said:
It has nothing to do with how fast the alcohol evaporates. 

It has to do with pure alcohol not being admitted inside the cell walls by osmosis.  If water is mixed with the alcohol, it tricks the cell wall into admitting it, therefore killing the cell. 70% has been shown to be a good mixture for sanitizing, and superior to higher concentrations.
RR




Quote:

bodhisatta said:
Alcohols
Although several alcohols have been shown to be effective antimicrobials, ethyl alcohol (ethanol, alcohol), isopropyl alcohol (isopropanol, propan-2-ol) and n -propanol (in particular in Europe) are the most widely used (337). Alcohols exhibit rapid broad-spectrum antimicrobial activity against vegetative bacteria (including mycobacteria), viruses, and fungi but are not sporicidal. They are, however, known to inhibit sporulation and spore germination (545), but this effect is reversible (513). Because of the lack of sporicidal activity, alcohols are not recommended for sterilization but are widely used for both hard-surface disinfection and skin antisepsis. Lower concentrations may also be used as preservatives and to potentiate the activity of other biocides. Many alcohol products include low levels of other biocides (in particular chlorhexidine), which remain on the skin following evaporation of the alcohol, or excipients (including emollients), which decrease the evaporation time of the alcohol and can significantly increase product efficacy (68). In general, isopropyl alcohol is considered slightly more efficacious against bacteria (95) and ethyl alcohol is more potent against viruses (259); however, this is dependent on the concentrations of both the active agent and the test microorganism. For example, isopropyl alcohol has greater lipophilic properties than ethyl alcohol and is less active against hydrophilic viruses (e.g., poliovirus) (259). Generally, the antimicrobial activity of alcohols is significantly lower at concentrations below 50% and is optimal in the 60 to 90% range.

iso is preferred to ethanol as a sanitizer because iso solubilizes fats better than ethanol does, hence being slightly more bactericidal

at 70% concentration it's hypotonic look up tonicity and osmolality to understand why this is important

Anyway ethanol and iso are interchangeable as sanitizer against non viral contaminants

You can go on if you would like I'm done here with this. You can be into facts or not.
90+ will inhibit the cell wall and not penetrate the interior as well thus not being as effective.

70% has a more effective osmotic pressure than 71-90

Evaporation is a secondary benefit

91+ % iso and ethanol are inferior sanitizer to 70 iso and ethanol

there's a reason hospitals and labs use 70% and it's not cost.

first and foremost the osmotic pressure of the 70% concentration is best the tonicity of this concentration means it will be able to denature the inside of a cell rather than surface proteins on the bacterial envelope. If a bacterial envelop is disabled the cell can come back to life later on since the inside of the cell hasn't been scrambled by the alcohol.
the added benefit is that 70% doesn't evaporate as fast this potentates it's effectiveness but is not the cause for the effectiveness. 







BEAUTIFUL  :takingnotes:

i used 91% to sterilize my syringe and more, might have a contam on my agar.



That wont sterilize your syringe, flaming till it glows red will. Iso only sanitizes you qouted all that but didnt read it.


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InvisibleDigit
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Re: pour agar TEK [Re: mrmazdarx9]
    #24331688 - 05/19/17 02:13 AM (3 years, 4 months ago)

Hi all, I've been lurking around for a while and I really appreciate the knowledge I've gotten here. So anyway, a while back I did a few MS PF jars with some old syringes from a friend. Most were shit but the Gulf Coast strain came out pretty good. Now I've cloned a nice fruit, grew it out on MEA in plates, made some LC from a couple plates and some grain masters with wedges. Things look great so far and some of my original plates have started pinning so I put a pin on agar. It's growing out nicely and I'm pretty surprised but so far I haven't had a single contam. :smile: Anyway I had such a good time with that project that I decided to try a few different spore syringes (fresh ones from SW) on agar. I made 5 plates with one drop from each syringe, and one or two plates from each has really aggressive rhizo growth and no obvious sectors. My question is should I further isolate or just go straight to grain or LC with the best plates and see what I get, then clone a nice fruit for further projects?
  Cheers


--------------------
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Believe nothing.
Think for yours3lf.


Edited by Digit (05/19/17 02:15 AM)


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Offlinejyphotog
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Re: pour agar TEK [Re: Digit]
    #24342289 - 05/23/17 09:28 AM (3 years, 3 months ago)



--------------------


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InvisiblebodhisattaM
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Re: pour agar TEK [Re: jyphotog]
    #24342394 - 05/23/17 10:33 AM (3 years, 3 months ago)

Terrible idea


--------------------
police violence is no accident
ACAB
There's two kinds of people. Those who have had a bad interaction with cops and those who haven't got their turn


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Offlinejyphotog
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Re: pour agar TEK [Re: bodhisatta]
    #24343028 - 05/23/17 03:31 PM (3 years, 3 months ago)

ok


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OfflineOrganic_Magic
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Re: pour agar TEK [Re: jyphotog]
    #24343831 - 05/23/17 07:41 PM (3 years, 3 months ago)

Quote:

jyphotog said:
Hey guys.

Can I get pre-made from Amazon or is that a bad idea?

https://www.amazon.com/Nutrient-Dehydrated-Sterile-Dishes-Cotton/dp/B00B799PDG/ref=sr_1_2?ie=UTF8&qid=1495545623&sr=8-2&keywords=petri+dish+with+agar





shits already contaminated in the stock photo :lmafo:


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Offlineloftymeat
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Re: pour agar TEK [Re: bodhisatta]
    #24370934 - 06/02/17 03:05 PM (3 years, 3 months ago)

Hi guys, new to mycology and have been reading all of this exelent tek from BOD over the last few weeks.  I looked through this agar tek and have searched the forums, but I still have a question about the wrapping of the dishes.  Do you wrap before or after inoculation, or both before and after?  I would probably pour and inoculate in the same session (after agar cools down), so that might affect this answer.

And if you wrap after inoculation, won't the wrap cuttoff any gas exchange? Is gas exchange not really necessary?

Any help would be appreciated, I'll keep searching for the answers.


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Offlineverum subsequentis
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Re: pour agar TEK [Re: loftymeat]
    #24371005 - 06/02/17 03:30 PM (3 years, 3 months ago)

If you pour in a still air box then you pour the agar and leave the petris in stacks to cool. Most folks cover the stacks with the sleeve they came in to allow them to cool inside the SAB. Don't wrap before inoculation unless your saving them for a while. I've left them in the sleeve for a few weeks before knocking them up with no wrap no problem.

    Inoc and then wrap. You want to wrap in para film or glad cling wrap. Both of these allow for gas exchange. Some people swear by para film and others by Glad. I use both but I believe Glad is cheaper, easier, and perfectly sufficient. Just by a roll and cut it into the right size pieces and you'll have enough to last you a while.
   
      I believe i read that bod doesn't wrap at all but that would make me nervous.

      also, don't underestimate the power of the search function. Click the search posts link and set it to Trusted cultivator and search you balls off. Almost everything you could ever need to know is there a thousand times over.

    Peace.


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Offlineverum subsequentis
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Re: pour agar TEK [Re: verum subsequentis]
    #24371038 - 06/02/17 03:42 PM (3 years, 3 months ago)

https://www.shroomery.org/forums/showflat.php/Number/14991978#14991978

That is the first link that comes up if you 1 click search post at the top right of the page 2. type "wrapping petri" in the words section 3 click Trusted Cultivator 4 select mushroom cultivation on the left. 5 hit enter.


          TONS of info is literally at your fingertips.


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Mushrooms, Mycology and Psychedelics >> Mushroom Cultivation

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* agar cultivation - easy! felixhigh 1,998 3 11/28/01 03:16 AM
by BBShrizz

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