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Offlinemyaltreality
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Some questions on Liquid Culture Preparation with a magnetic stir plate * 1
    #11810101 - 01/11/10 08:17 PM (11 years, 4 months ago)

When comparing the growth of a liquid culture prepared with organic honey that has been on a magnetic stirrer to a similar liquid culture of the same exact preparation that had been manually agitated via shaking twice daily, the one in constant motion exhibits long stringy growth as compared to the one that is manually agitated, which appears fluffy and wispy.

Is this common, or did I do something wrong?  How will it affect it's growth characteristics?  I just started a dozen 1/2Pint BRF cakes of each type, and labeled them appropriately to see if the physical characteristic exhibited changes the colonization rate of the cakes, or the overall fruiting mass on the cakes. 

Has anyone compared this prior?  From my reading of the forums and knowledge bases, a magnetic stir plate GREATLY increases the rate of colonization, but there were notes regarding the growth characteristics that I was able to easily find. 

Is there a measurable effect on mycelium growth on substrate when prepared with a magnetic stir plate, rather than letting the jars colonize more slowly?


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OfflineRogerRabbitM
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Re: Some questions on Liquid Culture Preparation with a magnetic stir plate [Re: myaltreality]
    #11811350 - 01/11/10 11:06 PM (11 years, 4 months ago)

Zap it with an eberbach for a second or two and then agitate for a week.  It should be fully done after that.  You don't want long strings of mycelium any more than you want blobs.  You want thousands of points of inoculation after you inject.
RR


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Offlinemyaltreality
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Re: Some questions on Liquid Culture Preparation with a magnetic stir plate [Re: RogerRabbit]
    #11812303 - 01/12/10 01:45 AM (11 years, 4 months ago)

Thanks for the reply, RR.  After a bit of quick googling and deductive reasoning, i'm lead to believe that what you're referencing is a lab grade blender, right?

I have a relatively inexpensive $80~ Hanna Instruments magnetic stirrer, without set RPM markings on it, I'd imagine that at 20% power the unit would spin at approx 200rpm, as the stirrer has a max of 1000rpm, and it appears to increase in speed at a constant rate as I adjust it's speed.

Is there an easy way to determine the proper speed at which I should be agitating the mixture?  It appears that 200rpm is the suggestion i'm finding in search results, what do you recommend?

Also, I found that by sucking up some of the mixture with a sterile syringe and squirting it forcefully back into the mixture, I can easily break up mycelium growth into thousands of smaller pieces.  Is it recommended to let the mycelium recover before inoculating any substrate, or is it safe to inoculate immediately after splitting it apart?

I apologize for bombarding you with more questions after reading your reply to my original post, but it's so much easier to get straight answers to some less than common questions, by directly inquiring with a trusted cultivator and long standing member of the forums. 

This forum has been incredibly rewarding in my thirst for knowledge, and everyone on here have been very helpful as I continue to make and learn from my mistakes and perfect my technique.  I spent many months lurking and reading posts by some of the more active and long standing members before creating an account.

Thank you again for your continued dedication and support to the forums!


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Offlinecalavera
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Re: Some questions on Liquid Culture Preparation with a magnetic stir plate [Re: myaltreality]
    #11812928 - 01/12/10 05:07 AM (11 years, 4 months ago)

hello.

i am into making lc's pretty much. some times i have succes like in heaven, and sometimes i get faliures like dont know what.

i use screws (that designed to drill in wood) insted of stirring rods. - with sharp edges, so it can stirr the lc, but also cut it up at higher speeds.

i only cut it up like twice. first after the agar wedge recovers,
and i can tell that the growth is only from there, no contamination,
so i cut it up, then stirring it only after before the injection, when i cut it up like hell:)

i figured out that making lc's is only constanlty good, from petri dishes. i think i have an experienced eye, and i could tell trich from cubes in an lc once, so i injected trich, my jars was full green in 2 days:D:D  i was making lc's from spores back than-

its good as well to make lc's from tissues, but its risky as well.
petri is the way to go INMHO.

i also use an automated injector (hose into lc, than just inject), wich works like a charm!


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Edited by calavera (01/12/10 05:08 AM)


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Invisiblefastfred
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Re: Some questions on Liquid Culture Preparation with a magnetic stir plate [Re: calavera]
    #11814639 - 01/12/10 02:34 PM (11 years, 4 months ago)

You can't go wrong with mag stirring.  I don't think the speed will matter much.

It won't increase the rate of colonization much if at all, but it will allow you to grow up the cultures much thicker.

It's all about the agitation increasing the dissolved oxygen in the liquid.  So as long as there is circulation in the culture you'll be getting the results you're looking for.  Just don't grow it up so thick you can't suck it up with a needle.


-FF


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