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Offlinemushpunx
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Re: Fully colonized grain Spawn to Liquid Culture [Re: scifipirate]
    #25391682 - 08/16/18 05:53 AM (5 years, 5 months ago)

I would advise against the GLC. You just can't tell how clean the grain spawn is, even minute amounts of bacteria will expand in the LC.

Start LC from clean cultures on agar. You can end up with more LC than you know what to do with, so just do it right.

Or you cab inoculate a grain master from the culture, you can go 1 to 10, 10 to 100 if you needed. Its just as fast.


But OP, if you have clean LC to begin with why do you need to make a new one anyways?


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Edited by mushpunx (08/16/18 05:56 AM)


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Offlinemushpunx
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Re: Fully colonized grain Spawn to Liquid Culture [Re: HiLIFE] * 1
    #25392109 - 08/16/18 10:46 AM (5 years, 5 months ago)

Quote:

HiLIFE said:
Quote:

scifipirate said:
Quote:

HiLIFE said: Pretty sure I can just break up the fully colonized grain spawn and fill it with water and then suck it back out and have a new LC right?



This is correct, go for it. Used to be a pretty standard way of making LCs, guess it's lost popularity...




So basically what I ended up doing was putting 60ml of sterilized LC into the jar, and then sucked up as much as I could back out and added back into the LC jar where it should grow to a new LC that i'm sure is clean. As soon as I get a flowhood i'll be doing g2g but until then. I feel like LC is just so easy though.





You mean 60ml sterilized LC *broth* I take it?

This was extremely unnecessary man. No need to use LC broth, you simply squirt water into the spawn jar and then suck it back out, that's it.
No need to grow a new culture.

But honestly, anything you get from going this route is most likely gunna be gross. Any minute amount of bacteria that was in the grain jar is going to expand in the LC broth.


If you absolutely must go this way using GLC , just squirt water in and pull it out.


No need for a flow hood for G2G at all!! Please tell me you are working with a still air box right now though? And are doing agar work?


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Offlinemushpunx
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Re: Fully colonized grain Spawn to Liquid Culture [Re: Caps McGee]
    #25393181 - 08/16/18 07:07 PM (5 years, 5 months ago)

Granted it will be hard in a tiny SAB, that's why as far as still boxes go the bigger the better.

But yea , of course. Anything that can be done in front of a flow hood can be done in an SAB, even spawn bags although you can't blow a plenum of sterile air in them before sealing.

In a still air box, contams will be settling/falling down , and stirred up by the air. That's why its important to make controlled, decisive hand motions, and to never move anything unsterile (such as your hands) over top any open sterile media. You don't want to leave sterile media open long either.

With a flow hood, sterile air is blown towards you in laminar flow pattern, which means if I were to drop spores above open media they would get trapped in the laminar flow and blow straight outwards instead of settling down into it. The difference here is that you never move anything unsterile *in front* of open media as contams might get blown in.

Flow hoods require better sterile technique than SABs. They are more about elbow room and convenience.


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