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Offlineeos666
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Does a LC jar need air exchange?
    #18148514 - 04/22/13 06:34 AM (10 years, 9 months ago)

ive read conflicting oppinions on LC jars - some say no air exchange needed, some say its needed.

Ive made some self healing lids with only a single hole sealed with silicone, will that be OK or does the jar need a 2nd hiole with tyvek or smth for air excahnge?


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Offlinemigraineur
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Re: Does a LC jar need air exchange? [Re: eos666]
    #18148542 - 04/22/13 06:47 AM (10 years, 9 months ago)

It's not essential to have gas exchange. The mycelium does need oxygen though. You can allow this by leaving a pocket of air in your jar when you make your liquid culture.

For example, if you use a pint jar to make a liquid culture then leave half of the space for air. You can inoculate your jar with a spore print or you can insert your spore syringe through the silicone and give it a squirt. This setup also makes the jar easy to shake and you won't get liquid on your breathable filter.

Once your jar is colonised then you can store it in the fridge for an extended period of time or you can put it into syringes and put those in the fridge.


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Invisiblewildernessjunkie
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Re: Does a LC jar need air exchange? [Re: migraineur]
    #18153468 - 04/23/13 12:28 AM (10 years, 9 months ago)

Leaving some "head space" in your container should leave plenty of oxygen for an LC. But to be clear, LC from MS is a really bad way to go.

My opinion is that if you can't do agar, then you shouldn't be messing with LC's. Though LC can be an extremely powerful tool if you can get it right.


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Offlinemigraineur
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Re: Does a LC jar need air exchange? [Re: wildernessjunkie]
    #18153764 - 04/23/13 03:40 AM (10 years, 9 months ago)

Quote:

wildernessjunkie said:
Leaving some "head space" in your container should leave plenty of oxygen for an LC. But to be clear, LC from MS is a really bad way to go.

My opinion is that if you can't do agar, then you shouldn't be messing with LC's. Though LC can be an extremely powerful tool if you can get it right.




Wildernessjunkie's statement has valid points but don't let it scare you. I have no problems making liquid cultures from prints or spore syringes but I am pretty good with my sterile technique. When you make a liquid culture you can test it on a jar or two first to see if it's clean. This can save you from inoculating heaps of jars only to have them fail from contamination.


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OfflinePussyFart
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Re: Does a LC jar need air exchange? [Re: migraineur]
    #18153932 - 04/23/13 06:24 AM (10 years, 9 months ago)

Quote:

migraineur said:
I have no problems making liquid cultures from prints or spore syringes but I am pretty good with my sterile technique.



You could have the best sterile technique, and still get a contaminated LC from spores.

The cap from which the spores came was fruited in the open air.

Therefore a print/syringe is never going to be 100% contam free.

An LC provides the perfect environment for contams to germinate and thrive.

This is why LCs from spores are frowned upon, even by the best.

Just :2cents:


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Offlineeos666
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Re: Does a LC jar need air exchange? [Re: migraineur]
    #18153977 - 04/23/13 07:01 AM (10 years, 9 months ago)

what do people do with agar anyway?

so far the LC jars ive prepared seem fine, they are only a few days old though.. making hem was no problem though using a PC, self healing lids and spore syringes


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OfflinePussyFart
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Re: Does a LC jar need air exchange? [Re: eos666]
    #18154009 - 04/23/13 07:27 AM (10 years, 9 months ago)

Quote:

eos666 said:
what do people do with agar anyway?



You can isolate single sets of genetics, transfer away from contams, store cultures for years, posssibly decades, and you can know for sure the inoculant  is 100% clean.

None of which can be done with LCs.


Edited by PussyFart (04/23/13 07:27 AM)


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Offlineeos666
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Re: Does a LC jar need air exchange? [Re: eos666]
    #18154217 - 04/23/13 08:48 AM (10 years, 9 months ago)

mh yea sounds useful . any "agar for dummies" out there?


Edited by eos666 (04/23/13 08:48 AM)


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OfflinePussyFart
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Re: Does a LC jar need air exchange? [Re: eos666]
    #18154275 - 04/23/13 09:07 AM (10 years, 9 months ago)



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InvisibleMyco_Militia
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Re: Does a LC jar need air exchange? [Re: PussyFart]
    #18155225 - 04/23/13 12:31 PM (10 years, 9 months ago)

^What he said.  I think the RR videos are a great start for anyone looking to grow their own.  The chapters are quite well organized and detailed in techniques and the reasoning behind them.


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Invisiblefastfred
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Re: Does a LC jar need air exchange? [Re: Myco_Militia]
    #18155798 - 04/23/13 02:35 PM (10 years, 9 months ago)

Quote:

This is why LCs from spores are frowned upon, even by the best.




LC from spores is fine.  Thousands of people do it all the time.

If you want to multiply a drop of spore solution into a quart of inoculate it works just fine.  Not everybody has the time, skills, equipment, or desire to isolate.

Doing a proper isolate takes a long time.  Many people prefer a shotgun method that will guarantee results rather than taking the time to try and find a decent isolate.


-FF


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OfflinePussyFart
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Re: Does a LC jar need air exchange? [Re: fastfred] * 1
    #18155843 - 04/23/13 02:44 PM (10 years, 9 months ago)

Using agar does not take more time, nor is there a need to isolate when using it.

It is used to visually ensure a clean culture, and if this means learning a new skill, so be it.

In the end you will save time and possibly money, by not having to shoot up and wait for test jars.

In my opinion, making an LC from an agar wedge, or even a grain jar is safer, and more reliable than starting it from spores.

But once you get to the stage of having agar and grain jars, reverting to LC is kinda pointless, when one can just G2G to expand the mycellium, with far less risk than messing with syringes.

I hate syringes, lol.


Edited by PussyFart (04/23/13 02:45 PM)


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Invisiblefastfred
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Re: Does a LC jar need air exchange? [Re: PussyFart]
    #18155907 - 04/23/13 02:55 PM (10 years, 9 months ago)

If you're going to use agar you might as well do it right.  That means picking isolates, transferring them to other plates or slants, growing them out, determining if they're good isolates, then going back to your best isolates for spawn production.  That takes a lot of time.

If you don't do that you run the real risk of picking a poor performer and wasting all your time and materials.

If you frequently use dirty syringes then what you suggest might be one way to salvage them, but you probably should just get better a better spore source.

Getting clean spores really isn't too difficult.  Randomly reducing your genetic diversity to one, or even a few, individuals is not a good way to go.


-FF


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OfflinePussyFart
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Re: Does a LC jar need air exchange? [Re: fastfred]
    #18156021 - 04/23/13 03:13 PM (10 years, 9 months ago)

Quote:

fastfred said:
If you're going to use agar you might as well do it right.  That means picking isolates, transferring them to other plates or slants, growing them out, determining if they're good isolates, then going back to your best isolates for spawn production.  That takes a lot of time.

If you don't do that you run the real risk of picking a poor performer and wasting all your time and materials.



I get the fact that you might as well, but the point is it's just not needed, just one or 2 transfers from MS/contams would still give you very descent results. Then clone from a nice fruit to isolate.

There is no need to isolate from spores if you do not want to, because that would take a lot of time.

Many people have had great success with just one  or 2 transfers from MS.(i can provide links if need be, or you can just search it)

Quote:

fastfred said:
Randomly reducing your genetic diversity to one, or even a few, individuals is not a good way to go.




In one breath you make it seem like it would take forever to isolate down to monocultures, and in the next breathe you make it seem like one or 2 transfers would be detrimental to performance because of the narrowed down genetics.

Confusing....

It usually takes 3-5 transfers from spores to even start to see sectoring/genetic diversity.

One or 2 transfers, to clean up the mycellium, would not affect much of anything.


Edited by PussyFart (04/23/13 03:15 PM)


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Invisiblefastfred
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Re: Does a LC jar need air exchange? [Re: PussyFart]
    #18156175 - 04/23/13 03:37 PM (10 years, 9 months ago)

Say you pick one or two individuals and they turn out to be duds?  You can certainly find examples of this happening in various threads here.  They might be non-potent or non-fruiters.

I've just never heard of having much problem with MS LCs.  People have been doing them without trouble almost since the beginning.  Putting a thousand or so spores into your LC only takes one drop.

If the spores solution is so poor that every drop is contaminated then you really should work on your spore prep methods or find another vendor.  There's about 12 drops/ml, so 120 drops per 10cc syringe.  If you're not getting at least 50% success rate then you must have over 60 contams in the syringe.  Yeast and other microbes can certainly sometimes multiply in the syringe and disperse.

I've only ever come across syringes that were either at least plenty clean enough, or so fucked I wouldn't even bother trying to isolate from them.

To each their own I guess.  But MS LC is the standard way of doing things.  Shake well and use only one drop, and you'll be fine.


-FF


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OfflinePussyFart
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Re: Does a LC jar need air exchange? [Re: fastfred]
    #18156204 - 04/23/13 03:44 PM (10 years, 9 months ago)

Quote:

fastfred said:
But MS LC is the standard way of doing things.



Not really nowadays, not on these forums.

LCs from spores has kinda been kicked to the side, even by TCs.

It seems agar and G2G transfers are just more popular.

Most likely due to the increased success rate.

But like you said, to each their own.


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Invisiblefastfred
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Re: Does a LC jar need air exchange? [Re: PussyFart]
    #18156403 - 04/23/13 04:35 PM (10 years, 9 months ago)

That's good to know.  I haven't been around in quite some time.

In never had more than perhaps 5% failure rate with MSI LCs, with the exception of one bad batch of yeast contaminated syringes.


-FF


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OfflineZombiewonka
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Re: Does a LC jar need air exchange? [Re: fastfred]
    #18190066 - 04/29/13 11:53 PM (10 years, 8 months ago)

Fast fred and notahacker are both correct.But now I love isolating on agar.Its a good skill set to master.


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Re: Does a LC jar need air exchange? [Re: eos666]
    #28138295 - 01/13/23 05:04 PM (1 year, 14 days ago)

Another option is to go from LC to lc. If you buy liquid culture from a reputable vendor, once you get down to your last CC or two of liquid culture you can inject that directly into a pack of room temperature, iso sterilized, Capri Sun Pacific cooler. Then cover the straw hole that you just injected into with micropore tape. Then leave it to sit somewhere warm for about 3 weeks. After the first week you want to give it a good shake once or twice a day.


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Re: Does a LC jar need air exchange? [Re: StarshineBlue] * 2
    #28138468 - 01/13/23 06:38 PM (1 year, 14 days ago)

:facepalm:


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Offlinerumfor69
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Re: Does a LC jar need air exchange? [Re: bakedbeings] * 1
    #28138721 - 01/13/23 09:18 PM (1 year, 14 days ago)

Quote:

bakedbeings said:
:facepalm:




:lol: what did I just read


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OfflineDERRAYLD
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Re: Does a LC jar need air exchange? [Re: StarshineBlue]
    #28139022 - 01/14/23 06:07 AM (1 year, 13 days ago)

Quote:

StarshineBlue said:
Another option is to go from LC to lc. If you buy liquid culture from a reputable vendor, once you get down to your last CC or two of liquid culture you can inject that directly into a pack of room temperature, iso sterilized, Capri Sun Pacific cooler. Then cover the straw hole that you just injected into with micropore tape. Then leave it to sit somewhere warm for about 3 weeks. After the first week you want to give it a good shake once or twice a day.




Go home reddit.... you're drunk


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