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OfflineWeebl8bob
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Re: A *beginner friendly* method for making Liquid Culture [Re: fastfred] * 1
    #5855911 - 07/13/06 08:21 PM (17 years, 6 months ago)

Lol yeah obviously nutrasweet isnt going to work.. but i figure that a nector would work seeing as honey is just regurgetated nector that has been collected by a bee..

And if it Doesnt work ill just take a peice of a cake that im fruiting and use that to make a LC with some good ol' table sugar (which i KNOW works)


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"You can turn your back on a person, but never turn your back on a drug, especially when its waving a razor sharp hunting knife in your eye." - Hunter S. Thompson


Edited by Weebl8bob (07/13/06 08:25 PM)


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Invisiblefastfred
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Re: A *beginner friendly* method for making Liquid Culture [Re: Weebl8bob] * 1
    #5856543 - 07/13/06 11:51 PM (17 years, 6 months ago)

Hrrrrmmmfffp.... I take back what I said. It was some other species that won't grow on fructose. P.C. apparently grows fine on it.

GL


-FF


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OfflineWeebl8bob
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Re: A *beginner friendly* method for making Liquid Culture [Re: fastfred] * 1
    #5856600 - 07/14/06 12:16 AM (17 years, 6 months ago)

YAY! ^.^ Ill tell you all how it goes... Injected spores three hours ago


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"You can turn your back on a person, but never turn your back on a drug, especially when its waving a razor sharp hunting knife in your eye." - Hunter S. Thompson


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Offlineajna
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Re: A *beginner friendly* method for making Liquid Culture [Re: Weebl8bob] * 1
    #5885812 - 07/21/06 09:15 PM (17 years, 6 months ago)

ok guys, can someone take a look at these photos and tell me if it looks like mycelium? it's now been 11 days in the incubator, i have this clumpy, strandy substance in the jar but it still doesn't look all that different to the 'honey sediment' i had at the start (see last page).

http://img.photobucket.com/albums/v192/voltaaa/myc1.jpg
http://img.photobucket.com/albums/v192/voltaaa/myc2.jpg

i'm inclined to leave it going still (if infact this is not myc.), just wondering if starting a new LC would be worthwhile or if perhaps it's just taking longer for some reason.

any tips would be great, thanks :smile:


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Edited by ajna (07/21/06 09:16 PM)


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OfflineWeebl8bob
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Re: A *beginner friendly* method for making Liquid Culture [Re: ajna] * 1
    #5885955 - 07/21/06 10:11 PM (17 years, 6 months ago)

OKAY! The table sugar jar is doing great! The agave nector.. well I threw that junk out!
Trying maple syrup jar next :-D gunna birth the sampling cake tomorrow or sunday


--------------------
"You can turn your back on a person, but never turn your back on a drug, especially when its waving a razor sharp hunting knife in your eye." - Hunter S. Thompson


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Invisiblefastfred
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Re: A *beginner friendly* method for making Liquid Culture [Re: Weebl8bob] * 1
    #5886304 - 07/22/06 12:40 AM (17 years, 6 months ago)

> ok guys, can someone take a look at these photos and tell me if it looks like mycelium?

I can't tell shit from those photos.

My advice for you is to gently shake and then let the jar sit for awhile. The precipitates will settle to the bottom, but myc will usually float around in the bottom third for quite some time. Only experience will help you determine if it's myc. But if it doesn't settle to the bottom it's myc.


-FF


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Offlineajna
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Re: A *beginner friendly* method for making Liquid Culture [Re: fastfred] * 1
    #5886507 - 07/22/06 02:29 AM (17 years, 6 months ago)

ok cheers fred


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Offlineghostofbmarley
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Re: A *beginner friendly* method for making Liquid Culture [Re: ajna] * 1
    #5887886 - 07/22/06 03:29 PM (17 years, 6 months ago)

How obvious would contamination be in a LC prepared in this manner?

For example, if the lid was slightly more than cracked for a second during the scrape...

thanks
-ghost


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Edited by ghostofbmarley (07/22/06 03:31 PM)


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Invisiblefastfred
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Re: A *beginner friendly* method for making Liquid Culture [Re: ghostofbmarley] * 1
    #5889312 - 07/22/06 11:31 PM (17 years, 6 months ago)

> How obvious would contamination be in a LC prepared in this manner?

Good question. A lot of people ask about this. People are sometimes confused by the term "cloudy".

Here are a couple pics of a malt/dextrose LC...





They're pics of the same test tubes. See if you can guess which one has bacterial contamination.


The one on the right is clearly contaminated with either bacteria or yeast. The one on the left looks a little different than the center one, but that's just a trick of the light. If you look carefully through the solution you can see through it clearly. There is another row of tubes behind those and if you look closely you can see that on the left two tubes you can see that the image of what's behind them is clear and mostly in focus. On the test tube on the right, it's cloudy, fuzzy, indistinct, hazy, and out of focus.

That's what's meant by "cloudy".

Hopefully this will help people with their questions about LC contamination.


-FF


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It drinks the alcohol and abstains from the weed or else it gets the hose again. -Chemy

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Offlineghostofbmarley
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Re: A *beginner friendly* method for making Liquid Culture [Re: fastfred] * 1
    #5889887 - 07/23/06 08:09 AM (17 years, 6 months ago)

Thank you FF.  I can see that it would be more obvious in that type of LC.  I am not sure if the determination would be as easy in a honey-based LC - but I am an inexperienced researcher.  Opinion there?

I ended up checking the contam forum last night, and there, an oft-reached consensus was it may not be obvious until applying the LC to a test substrate jar...  Comments?

Thanks,
-ghost :psychsplit:


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Offlineabsinthoman
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Re: A *beginner friendly* method for making Liquid Culture [Re: ghostofbmarley] * 1
    #5890189 - 07/23/06 11:00 AM (17 years, 6 months ago)

Hi everybody, could somebody help me. I bought 3 spores syringes (taz, mexi-cube and strophoria)from Ralph Canada. I inoculated rye jars and only the strophoria developped a mycelium with contams so I decided to make an LC. I inoculate a taz LC of 250ml water/1tps of honey 2 days ago. I can't see any sign of grow yet??How long should I wait before concluding that my spores were dead???


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Offlineabsinthoman
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Re: A *beginner friendly* method for making Liquid Culture [Re: absinthoman] * 1
    #5890534 - 07/23/06 01:19 PM (17 years, 6 months ago)

anybody???


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Invisibleesdfsfd
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Re: A *beginner friendly* method for making Liquid Culture [Re: absinthoman] * 1
    #5890671 - 07/23/06 02:05 PM (17 years, 6 months ago)

IME if you leave the LC for long enough and it is contaminated, you will eventually see spores floating at the top of the LC.

'the strophoria developped a mycelium with contams so I decided to make an LC'
if you made an LC with contaminated mycelium you have failed.


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Invisiblefastfred
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Re: A *beginner friendly* method for making Liquid Culture [Re: esdfsfd] * 1
    #5891384 - 07/23/06 04:36 PM (17 years, 6 months ago)

> I am not sure if the determination would be as easy in a honey-based LC

I don't use honey so I can't say for sure. But with any media it's easy to get confused with color, darkness, and precipitates. You need to ignore those and look at the *clarity* of the solution. Try holding up some newsprint or something behind the mixture. If the solution makes the print fuzzy you almost certainly have bacterial or yeast contamination. Bacteria and yeast are single celled organisms, you can't see them in solution except for the cloudiness. I would think that honey would be just as easy to tell as anything else.


> an oft-reached consensus was it may not be obvious until applying the LC to a test substrate jar... Comments?

Fungal contaminants can be both easier or harder to spot. Sometimes they float, or are an obvious color. Bacteria and yeast should be easy to spot by the cloudiness. Having a control jar is VERY helpful in determining if you have bacterial contams. As you can see from the pic, it might be hard to spot by itself, but next to an uncontaminated sample the difference becomes obvious.

There are species of bacteria that cling to things or clump together, those would not be spotted by the clarity check. But most bacteria will float around freely in solution and cause cloudiness. Of course a lot of things can slip past your notice or be hard to detect, but the majority of the time you should be able to tell if the LC is contaminated.


-FF


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Offlineabsinthoman
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Re: A *beginner friendly* method for making Liquid Culture [Re: esdfsfd] * 1
    #5891555 - 07/23/06 05:26 PM (17 years, 6 months ago)

'the strophoria developped a mycelium with contams so I decided to make an LC'
if you made an LC with contaminated mycelium you have failed.



I made my LC with my taz spores. 5cc. And my question was more how long it usually take to see the cottony like mycelium in the solution??


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Invisiblefastfred
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Re: A *beginner friendly* method for making Liquid Culture [Re: absinthoman] * 1
    #5891604 - 07/23/06 05:41 PM (17 years, 6 months ago)

4-5 days


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OfflineSenselessRebel
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Re: A *beginner friendly* method for making Liquid Culture [Re: fastfred] * 1
    #5899803 - 07/25/06 09:35 PM (17 years, 6 months ago)

hey otto,
amazing tek!!! im gonna do this tomarrow!!
but i have a question??? would fresh honey from my bee hives be any better for the LC then store bought honey??? i know the honey from the store is made from bee's all the same but just a thought???
i donno may just be a pot head:)
still doing this tomarrow with some honey i got in my cabinet
i think imma try a few jars :smile:
wish me luck and im happy i came apon this tek!
thanks otto
-SenselessRebel


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Offline_OttO_
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Re: A *beginner friendly* method for making Liquid Culture [Re: SenselessRebel] * 1
    #5904547 - 07/27/06 07:32 AM (17 years, 6 months ago)

Well dude, generally, the honey you get in the jars on the shelf at the shop - is simply honey from a hive, filtered and bottled.

So, no - I don't think it will be any better using fresh honey from a bee hive. Just go with the store bought stuff and you will be fine.


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OfflineSenselessRebel
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Re: A *beginner friendly* method for making Liquid Culture [Re: _OttO_] * 1
    #5922160 - 08/01/06 03:11 PM (17 years, 5 months ago)

hahha yea figured.
thanks soo much for responding otto. i was wondering if you could take a look (if you had time) at my jars that i did useing this tek
i don't know if they are going well or wether they are contamed. if you have time please take a look
thanks again otto
heres the link
http://www.shroomery.org/forums/showflat.php?Cat=0&Number=5922068&page=0&vc=1

-SenselessRebel


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OfflineCepheus
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Re: A *beginner friendly* method for making Liquid Culture [Re: _OttO_] * 1
    #5922913 - 08/01/06 07:34 PM (17 years, 5 months ago)

I've been thinking about trying a LC. Thanks for this :laugh:


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