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OfflineAzmodeus
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Isolating a strain on agar * 1
    #1313859 - 02/17/03 09:59 AM (21 years, 1 month ago)

So i made up my first ever agar last night and innoculated with gulf coast spores.  I want to do some azure agar, but thought it best to practice a little first.  So before the agar reaches the sides i need to locate the healthiest part of the agar in order to transfer and isolate it right?  Well im just wondering what the best parts of mycelium to isolate are?  Is it the rhizo growth, fuzzy growth, quickest...?  And advice on the next steps, or techniques that worked for you would also be much appreciated...

Thanks :smile:


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InvisibleJoshua
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Re: Isolating a strain on agar [Re: Azmodeus]
    #1313886 - 02/17/03 10:08 AM (21 years, 1 month ago)

It is generally accepted that the fast rhizomorphic growth should be isolated.

Joshua


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OfflineAzmodeus
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Re: Isolating a strain on agar [Re: Joshua]
    #1313988 - 02/17/03 10:39 AM (21 years, 1 month ago)

Good to know!!! :smile:
This mycelium that i am isolating.....does it effect fruit size and body? or is it just for mycelial traits?  what is involved with "domesticating" a strain?.....



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OfflineSeussA
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Re: Isolating a strain on agar [Re: Azmodeus] * 1
    #1314204 - 02/17/03 11:48 AM (21 years, 1 month ago)

The purpose of isolating mycelium is to get a single genetic crossing from two spores rather than a racemic. An isolate will fruit at the same time. The fruit will be close to the same size and strength. Of course, you might end up isolating a nonfruiting mycelium, in which case you will not get any mushrooms at all.

To domesticate a strain, you have to grow the fruit to maturity and collect spores, usually from the best performing fruits. In this case, you want a multispore innoculation rather than an isolate to generate genetic diversity. After many generations, the fruit you are growing will become domesticated (used to growing in the parameters you have selected). This process takes a lot of time and effort.


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OfflineAzmodeus
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Re: Isolating a strain on agar [Re: Seuss]
    #1315909 - 02/18/03 04:19 AM (21 years, 1 month ago)

O.K, thanks!

What are the chances of isolating a "non-fruiting" mycelium?

And with domesticating, all you do is fruit, take the spores from the most charismatic fruit, and grow again in the exact same perameters.....how many times must one do this for it to be considered "domesticated"?


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OfflineMycolaureat
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Re: Isolating a strain on agar [Re: Azmodeus]
    #1318882 - 02/19/03 07:48 AM (21 years, 1 month ago)

hey Azmo
If your prints was not taken from the wild, it is already domesticated.



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OfflineAzmodeus
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Re: Isolating a strain on agar [Re: Azmodeus]
    #1321876 - 02/20/03 09:19 AM (21 years, 1 month ago)

Can anyone answer my questions?
:confused:


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InvisibleJoshua
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Re: Isolating a strain on agar [Re: Azmodeus]
    #1322033 - 02/20/03 10:17 AM (21 years, 1 month ago)

does it effect fruit size and body?

It may...but on a complicated level. I think if you have more pins as rhizomorphic growth usually has you will have many small mushrooms. If you have a few pins such as cottony growth usually has you will have a few large shrooms. Other than that loose correlation, I don't think I have seen enough data to give you an informed answer.

At this point I see you have questions that can bee easily answered by reading a book. Try The Mushroom Cultivator or Growing Gourmet and Medicinal Mushrooms both by Paul Stamets.

Educate yourself.

Joshua


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InvisibleJoshua
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Re: Isolating a strain on agar [Re: Azmodeus]
    #1322042 - 02/20/03 10:19 AM (21 years, 1 month ago)

A domesticated strain is a strain that performs well in a domestic atmosphere. You will have to see how well your isolation fruits.

IMO...PR are a non-domesticated strain. They do not follow domestic cultivation parameters as well as other more domesticated strains such as B+.

Joshua


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The Shroomery Bookstore

Great books for inquiring minds!

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Offlinehsalf
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Re: Isolating a strain on agar [Re: Azmodeus]
    #1322857 - 02/20/03 03:59 PM (21 years, 1 month ago)

2 out of 3

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Invisibledeanofmean
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Re: Isolating a strain on agar [Re: Joshua]
    #1323080 - 02/20/03 05:12 PM (21 years, 1 month ago)

Quote:


IMO...PR are a non-domesticated strain. They do not follow domestic cultivation parameters as well as other more domesticated strains such as B+.

Joshua




if it were, it might be as potent as B+ . lol

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InvisibleJoshua
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Re: Isolating a strain on agar [Re: deanofmean]
    #1324511 - 02/21/03 09:19 AM (21 years, 1 month ago)

Who says it is not?

Joshua


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OfflineRaadt
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Re: Isolating a strain on agar [Re: Joshua]
    #1324526 - 02/21/03 09:26 AM (21 years, 1 month ago)

but vendor hype of course! :wink:


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Raadt

-- The information I provide is only information from readings, growing of gourmet mushrooms, and second hand stories--

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OfflineAzmodeus
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Re: Isolating a strain on agar [Re: Raadt]
    #1324550 - 02/21/03 09:32 AM (21 years, 1 month ago)

I thought that B+ was known for its lack of potency?! :confused:
At least i always hear of people eating an 1/8 to get a lv. 3 trip....

Oh, and what are the chances of isolating a non-fruiting mycelium?  slim enough for me not to worry?


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"Know your Body - Know your Mind - Know your Substance - Know your Source.

Lest we forget. "

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OfflineRaadt
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Re: Isolating a strain on agar [Re: Azmodeus]
    #1324705 - 02/21/03 10:53 AM (21 years, 1 month ago)

with cubensis, i would say yes - very slim... this is simply a mushroom which is quite determined to reproduce.


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Raadt

-- The information I provide is only information from readings, growing of gourmet mushrooms, and second hand stories--

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Invisibledeanofmean
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Re: Isolating a strain on agar [Re: Azmodeus]
    #1324861 - 02/21/03 12:07 PM (21 years, 1 month ago)

Quote:

I thought that B+ was known for its lack of potency?! :confused:
At least i always hear of people eating an 1/8 to get a lv. 3 trip....




this was my point .i think B+ may be an example of a strain being over domesticated .i would hate to see that happen to a wonderful strain like PR ! :tongue: 
sorry about the confusion, i thought that previous statement was pretty obvious, but i guess it weren't .

Edited by deanofmean (02/21/03 12:21 PM)

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Offlinehsalf
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Re: Isolating a strain on agar [Re: Azmodeus]
    #1325177 - 02/21/03 04:02 PM (21 years, 1 month ago)

2 out of 3
1 out of 2 if you don't isolate the ovbious "cottony" growth. Half the rhizos will be of the non-fruiting variety.
Isolate a few times and then test a few rhizo mycellium for fruiting ability.

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InvisibleZen Peddler
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Re: Isolating a strain on agar [Re: deanofmean]
    #1325284 - 02/21/03 05:07 PM (21 years, 1 month ago)

it weren't?


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Invisibledeanofmean
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Re: Isolating a strain on agar [Re: Zen Peddler]
    #1325358 - 02/21/03 06:11 PM (21 years, 1 month ago)

to me it was :smirk: 

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OfflineRaadt
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Re: Isolating a strain on agar [Re: hsalf]
    #1325503 - 02/21/03 07:47 PM (21 years, 1 month ago)

uh, have you ever isolated a strain? tormentose as hell still fruits for me np.


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Raadt

-- The information I provide is only information from readings, growing of gourmet mushrooms, and second hand stories--

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InvisibleZen Peddler
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Re: Isolating a strain on agar [Re: Raadt]
    #1326695 - 02/22/03 01:10 PM (21 years, 1 month ago)

Exactly - with cubies sometimes the most cottony shity mycelia pins like mad when the plate is full colonised. You never know until you let it pin invitro (unintentional rhyme!)


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Offlinehsalf
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Re: Isolating a strain on agar [Re: Raadt]
    #1329169 - 02/23/03 02:59 PM (21 years, 1 month ago)

With all 'our' cherished mushies? or just cubies? funny how Stamets would leave that out
nope. just an old reclusive bookworm trying to help out on a good cause. I'd never do anything illegal. now when I leave the states I may try it out. goddamn system already stole my "life(lOve)" once. She's tradin' chemicals w/the cap'in now, I hear. Fuck the 'drug' war.

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Offlinehsalf
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Re: Isolating a strain on agar [Re: Raadt]
    #1329258 - 02/23/03 03:47 PM (21 years, 1 month ago)

as an after thought do you(or anyone here) know of any tests/experiments done as to the growth rates/yeild of fruiting cotton picking(tomentose) mycellium?
I'm willing to bet, a small amount, that it prob won't be as great as the other.

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OfflineAzmodeus
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Re: Isolating a strain on agar [Re: hsalf]
    #1330695 - 02/24/03 05:01 AM (21 years, 1 month ago)

So cottony growth is better to isolate?! Or rhizo with some cottony growth as well? :confused:


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Lest we forget. "

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InvisibleG a n j a
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Re: Isolating a strain on agar [Re: Azmodeus]
    #1330729 - 02/24/03 05:18 AM (21 years, 1 month ago)

i would go with rhizos.They seem to spread out and fill whatever medium faster.


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OfflineRaadt
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Re: Isolating a strain on agar [Re: Azmodeus]
    #1330842 - 02/24/03 06:09 AM (21 years, 1 month ago)

rhizo, but cottony with cubes doesn't seem to matter. Rhizo are always best though, in my opinion. (They are faster growing also)


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Raadt

-- The information I provide is only information from readings, growing of gourmet mushrooms, and second hand stories--

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OfflineAzmodeus
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Re: Isolating a strain on agar [Re: Raadt]
    #1330935 - 02/24/03 06:48 AM (21 years, 1 month ago)

K, thanks guys...
Its been around 5 days, im gonna check my agar and see if its working...


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Offlinehsalf
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Re: Isolating a strain on agar [Re: Azmodeus]
    #1331711 - 02/24/03 12:27 PM (21 years, 1 month ago)

when you get to those azures make sure to ONLY use rhizo

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OfflineRaadt
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Re: Isolating a strain on agar [Re: hsalf]
    #1331775 - 02/24/03 12:46 PM (21 years, 1 month ago)

my azure culture isn't very rhizo - it's very thick satiny mycellium, and only the very tips have anything that look like rhizomorphs. Where did you hear that azure cultures have to be pure rhizo?


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Raadt

-- The information I provide is only information from readings, growing of gourmet mushrooms, and second hand stories--

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InvisibleJoshua
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Re: Isolating a strain on agar [Re: Raadt]
    #1332830 - 02/24/03 08:48 PM (21 years, 1 month ago)

It sounds like you are being azurescending.

The isolate I fruited from in my outdoor bed was cottony. I think all of this talk of rhizomorphs is interesting. I recently transfered a rhizomorphic isolation of USVI to millet, once the millet was fully colonised I then transfered a few seeds back to agar. the resulting mycelium was extremely thick and cottony.

So I thought...very interesting.

Joshua


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Offlinecanid
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Re: Isolating a strain on agar [Re: Joshua]
    #1332842 - 02/24/03 08:55 PM (21 years, 1 month ago)

josh, i have found it interesting how many are of the mind that linear mycelium is worthless and cultures must be agressively rizhomorphic to be good fruiters. people seem to forget that this is but one element that will determine how a strain fairs in propagation.


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



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Offlinezeronio
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Re: Isolating a strain on agar [Re: canid]
    #1332849 - 02/24/03 09:04 PM (21 years, 1 month ago)

Some mushrooms are cottony on agar and become rhizomorphic on other substrates. The reason for forming rhizomorphes could be in substrate - mushroom tries to cover a bigger area to get enough nutrients.

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OfflineRaadt
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Re: Isolating a strain on agar [Re: Joshua]
    #1333167 - 02/25/03 02:47 AM (21 years, 1 month ago)

I can't make a bed, since it wont fruit here. But I have a culture, that I isolated from a print - no idea if it's fruitable. So I was just wondering if that was useless, or if it had to be a rhizo culture, because pictures i've seen of azure cultures have not been overly rhizo either.


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Raadt

-- The information I provide is only information from readings, growing of gourmet mushrooms, and second hand stories--

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Anonymous

Re: Isolating a strain on agar [Re: Azmodeus]
    #1333375 - 02/25/03 04:34 AM (21 years, 1 month ago)

hey... i think TMC says that 3/4 pure cultures will not produce fruits... so you need to isolate quite a few, then test them, then use them when you know they're good. if you want a pure fruiting culture, why not clone a fruitbody grown from simple multi-spore inoculation?

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OfflineAnnoA
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Re: Isolating a strain on agar [Re: ]
    #1333408 - 02/25/03 04:45 AM (21 years, 1 month ago)

>why not clone a fruitbody grown from simple multi-spore inoculation?

That?s the preferred and easiest way, yes.

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Offlinethe spiral
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Re: Isolating a strain on agar [Re: ]
    #1334347 - 02/25/03 09:53 AM (21 years, 1 month ago)

Quote:

why not clone a fruitbody grown from simple multi-spore inoculation?





that also has the added benefit that you can use peroxidated agar, or not use agar at all.


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InvisibleZen Peddler
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Re: Isolating a strain on agar [Re: Raadt]
    #1334426 - 02/25/03 10:23 AM (21 years, 1 month ago)

I generally find that woodlovers like ps.cyanescens/azurescens/subaeruginosa tend quickly towards rhizomorphic mycelia, and even if it is cottony, its soon rhizomorphic when used to innoculate wood-based substrates.
Although Id prefer rhizos and usually stear towards them on agar, I also go on speed and primordia formation with all my mushrooms.


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Edited by Zen Peddler (02/25/03 10:24 AM)

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Offlinehsalf
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Re: Isolating a strain on agar [Re: Raadt]
    #1335069 - 02/25/03 01:49 PM (21 years, 1 month ago)

GGMM: "myc is cottony at '1st', soon silky rhizo." "the bases of the stems radiate clusters of thick white rhizos." Going by this I guess I'd wait to make sure there is a change.

Quote:

Rhizo are always best though, in my opinion. (They are faster growing also) 


 
somewhere in there is an answer to your question. :smirk:

rhizos are also best in stamet's, chilton's, McKenna bro's and the authors of the many textbooks on mycology here. So why tell someone to go at it half assed just 'cause they'll get some results?
why take the train when you can...
...walk. :confused:

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OfflineAzmodeus
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Re: Isolating a strain on agar [Re: hsalf]
    #1336944 - 02/26/03 05:40 AM (21 years, 1 month ago)

I check my petri....only two cus they are stacked...i have 8 in total. I dont see any sign of mycelial growth...but no contam either...
Its been 5 or 6 days....is it still too early?!


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Lest we forget. "

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OfflineRaadt
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Re: Isolating a strain on agar [Re: hsalf]
    #1337017 - 02/26/03 06:14 AM (21 years, 1 month ago)

well, my friends GT isolate that he's been using for a year - is NOT rhizo, it's extremely linear - yet it's the best fruiting isolate i've seen. Some clones even don't take as rhizo growth, but linear - I think you're just trying to stay away from tormentose growth. (overly fluffy/dense/slow)


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OfflineMycolaureat
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Re: Isolating a strain on agar [Re: Azmodeus]
    #1337022 - 02/26/03 06:15 AM (21 years, 1 month ago)

Have you had some peroxide to your agar ?

With peroxide agar, sometime it takes me more than 8 days to see the first sign of growth.
But normal PDY agar show mycelium growth after 4-6 days.


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OfflineAzmodeus
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Re: Isolating a strain on agar [Re: Mycolaureat]
    #1337072 - 02/26/03 06:51 AM (21 years, 1 month ago)

No i did not add peroxide this time......
I'll wait a day or two then check all teh petris... :crazy:


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Lest we forget. "

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Anonymous

Re: Isolating a strain on agar [Re: Azmodeus]
    #1342393 - 02/28/03 07:12 AM (21 years, 1 month ago)

Okay guys this is a lil off topic but it falls under agar isolation. My friends isolates have a bacterial contam. The mycelium is growin rather well with the contam all around it. I took some straight 3% H2O2 and wrinsed of the plates. It seemed to have worked rather well. The isolates are spreading faster now. And the bacteria has dwindled away. Now time for next sectoring.
Has any one had any experience with this same situation? the man also has some agar made up with h2o2 in it but it has solidified what can he do to liquify it with out desturbing the h2o2 in it?
peace

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