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Offlinexaxphaanes
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QUESTION redboy lc
    #6343358 - 12/07/06 03:44 AM (17 years, 6 months ago)

ok i have done lc's like maybe 20 times and this one is just being super stubborn ok i have made a lc with 4% karo to a small jar of water using the 4ml karo and 96 ml water only thing i changed was tripled it to make a larger jar and it has been 5-6 days and not one strand of myc is in that jar.i have made 2 other lc's south americans and burmese yangoon and they have already 14% colonized what gives is redboy really this slow ???


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InvisibleBlutjager
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Re: QUESTION redboy lc [Re: xaxphaanes]
    #6343398 - 12/07/06 04:49 AM (17 years, 6 months ago)

Hmmm,I don't know what to tell you but I can tell you that it happens to me every now and than,Sometimes I will make a ton of different lc's at once and many times there is one that just plain lags behind

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OfflineRogerRabbitM
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Re: QUESTION redboy lc [Re: Blutjager]
    #6344136 - 12/07/06 11:08 AM (17 years, 6 months ago)

Throwing spores into honey or karo has to be the worst possible way to make a liquid culture and I'll be glad when this current fad passes and folks can get back to established techniques. Just inoculate some grains or brf in the standard way and you'll be miles ahead.

For those who wish to make LC for inoculation, here's a way to get 100 syringes full of inoculant in two weeks: 1) Inoculate a quart jar of rye berries or wbs with your spores. Upon full colonization, shake the jar to loosen each individual kernel. Be sure to give it the smell test to make sure it smells like fresh mushrooms. 2) PC a quart of distilled water in a jar with a filter disk or tyvek, etc. In a glovebox or in front of a flowhood, pour 2/3 of the quart jar of sterilized water into your jar of shaken grains. 3) Shake well and then pour the now mycelium rich water back into whatever was left of your jar of sterilized water. Use the jar lid to hold back the grains themselves from pouring out. You now have a full quart of highly concentrated mycelium water that can then be used as is, or diluted two or three times again, and used to fill syringes. Because the grains were shaken first, the mycelium is ripped into shreds and will be sucked through even small needles easily, although larger needles always work better for this.

Remember, mycelium grows poorly in water unless under constant stir which oxygenates it. With grains, they colonize in two weeks or less because oxygen is throughout the jar in the spaces between the kernels.
RR


--------------------
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semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
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Invisiblecreamcorn
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Re: QUESTION redboy lc [Re: RogerRabbit]
    #6344185 - 12/07/06 11:23 AM (17 years, 6 months ago)

awww come on now don't be so harsh on our beloved LCs. 

they're a trade-off, you have to be willing to accept a less than perfect success rate in exchange for not having to deal with strict sterile procedures.  there's a lot of value in that though, and its shortsighted to write them off because its less than perfect.  if you had people following the procedure you mention on a regular basis, i promise there would be just as many "why didn't it work" questions.  you only see lots of problematic posts about LCs because LCs are so popular.  im sure if folks posted every time they had a successful LC the servers would catch fire and burn to the ground. :wink:

you make perfectly valid points, but established procedures are good for experienced pros with proper equipment - so that's easy for somebody like you to say!  most around these parts have neither experience or proper equipment... things like LC aren't just a fad and its not going to go away any time soon... get used to it rather than bash it. :wink:

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InvisibleGanja420Boy
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Re: *DELETED* [Re: RogerRabbit]
    #6344248 - 12/07/06 11:41 AM (17 years, 6 months ago)

Post deleted by Ganja420Boy

Reason for deletion: Old Post


Edited by Ganja420Boy (11/12/08 09:20 AM)

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Offlinetotal
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Re: QUESTION redboy lc [Re: creamcorn]
    #6344250 - 12/07/06 11:41 AM (17 years, 6 months ago)

Quote:

creamcorn said:
awww come on now don't be so harsh on our beloved LCs. 

they're a trade-off, you have to be willing to accept a less than perfect success rate in exchange for not having to deal with strict sterile procedures.  there's a lot of value in that though, and its shortsighted to write them off because its less than perfect.  if you had people following the procedure you mention on a regular basis, i promise there would be just as many "why didn't it work" questions.  you only see lots of problematic posts about LCs because LCs are so popular.  im sure if folks posted every time they had a successful LC the servers would catch fire and burn to the ground. :wink:

you make perfectly valid points, but established procedures are good for experienced pros with proper equipment - so that's easy for somebody like you to say!  most around these parts have neither experience or proper equipment... things like LC aren't just a fad and its not going to go away any time soon... get used to it rather than bash it. :wink:




*nods*
not everyone has a flowhood...some of us still work in our bathroom

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Invisiblemusher_420
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Re: QUESTION redboy lc [Re: RogerRabbit]
    #6344260 - 12/07/06 11:45 AM (17 years, 6 months ago)

I haven't had great success from multispore inoculation into an LC jar. But I have found great results from germinating on agar then transferring a wedge to the LC jar. I find LC's while not as fast as grain to grain transfers still preform very well and it's easier to work with LCs when one only has a glove box for sterile work.

I find it's handy for storing cultures too.


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InvisibleBlutjager
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Re: QUESTION redboy lc [Re: Ganja420Boy]
    #6344556 - 12/07/06 01:05 PM (17 years, 6 months ago)

Quote:

Ganja420Boy said:

Does this mean it dont work or just a very low success rate?
I just did this yesterday sure hope I didnt waste my valuable spores on something so crapy :sad:




It will more than likely work just fine,don't worry just leave it alone for a week and than go look at it  :thumbup: :thumbup:

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OfflineRogerRabbitM
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Re: QUESTION redboy lc [Re: musher_420]
    #6344609 - 12/07/06 01:25 PM (17 years, 6 months ago)

A glovebox is not advanced equipment. One can be made free with stuff around the house, so there's no need for people to work in open air or fecal and mold infested bathrooms and kitchens. There is also no way to generate 100 full syringes of liquid mycelium in very small pieces within two weeks from injecting spores into honey or karo.

I'm sure if people blindly used the grain method I described above, many would still have problems too, because they're inserting unproved and possibly contaminated spore solution into grains, which also isn't proper mycological technique.

However, agar can easily be poured in a glovebox, and the petri dishes used to isolate healthy mushroom mycelium away from any contaminants in less time than it takes to even know if a karo LC is contaminated or not. That agar wedge can then be used to inoculate the quart(or pint) jar of grains, that can then be turned into at least 100 syringes, or used for g2g transfer to get ten jars the old standard way.

In addition, after the water is poured out of the grain jar, it can be placed back on the shelf to re-colonize, and the process can be repeated again.

I just strongly feel new growers are doing themselves a disservice when they simply inject spores into honey water and then sit there waiting for something to happen. At the very least, inject directly to grains so you can visually inspect the process. Perhaps Agar's lids with injection ports will help those who have never used grains to get off to a good start.
RR


--------------------
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semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
Thomas Edison

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OfflineGrogan
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Re: QUESTION redboy lc [Re: RogerRabbit]
    #6344633 - 12/07/06 01:34 PM (17 years, 6 months ago)

I advise using honey, and pressure cooking your lc.. dont keep it at like 80+ degrees either..


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Offlineetam
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Re: QUESTION redboy lc [Re: RogerRabbit]
    #6344702 - 12/07/06 02:05 PM (17 years, 6 months ago)

Quote:

RogerRabbit said:
I just strongly feel new growers are doing themselves a disservice when they simply inject spores into honey water and then sit there waiting for something to happen. At the very least, inject directly to grains so you can visually inspect the process. Perhaps Agar's lids with injection ports will help those who have never used grains to get off to a good start.
RR




I'm glad you are trying to do new growers a service by telling them to avoid LC's, but the fact is that LC's work. One of a newb's greatest fears is running out of spore solution and not having anymore leftover when their first PF attempt fails. Having an LC full of inoculant solves that problem completely. They can store a few cc's of the original spore solution away in the refrigerator while using the LC to perfect some of their new skills.

Not all of us are into cloning. For those of us who want to take this hobby as far as we can, you are right. I'm sure I will eventually build a glove box and start doing some agar transfers to grain. But in the mean time, we have to learn somewhere, and LC's are great for that. I think LC's are a logical step in the learning process before one jumps from straight spores w/ the PF tek to cloning with agar.

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OfflineGrogan
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Re: QUESTION redboy lc [Re: etam]
    #6344744 - 12/07/06 02:20 PM (17 years, 6 months ago)

Yes they are etam, Lcs are a great thing.. Belive me I will do agar but until then I use LC's


--------------------
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OfflineRogerRabbitM
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Re: QUESTION redboy lc [Re: etam]
    #6344748 - 12/07/06 02:21 PM (17 years, 6 months ago)

You might want to read more than the last paragraph. I described a method to generate ten times as much LC in a shorter period of time without contamination. I'm not against liquid culture. I use it all the time. I'm simply dead against the idea of squirting a syringe into a bottle of karo/water. I insist on having at least some verification my LC is good before I inject a bunch of jars.

The analogy of a new growers greatest fear doesn't hold water. If his pf jars contaminated due to contaminated spores/syringe, it does no good to have even ten gallons of contaminated LC on hand. If they contaminated due to his sloppy procedure, he's likely to have used the same sloppy procedure making an LC.

In my posts, I always try to show a way to have the greatest possible success rate, not necessarily the cheapest, easiest or laziest method.
RR


--------------------
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semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
Thomas Edison

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Offlineetam
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Re: QUESTION redboy lc [Re: RogerRabbit]
    #6344768 - 12/07/06 02:26 PM (17 years, 6 months ago)

Quote:

RogerRabbit said:
You might want to read more than the last paragraph. I described a method to generate ten times as much LC in a shorter period of time without contamination. I'm not against liquid culture. I use it all the time. I'm simply dead against the idea of squirting a syringe into a bottle of karo/water. I insist on having at least some verification my LC is good before I inject a bunch of jars.

The analogy of a new growers greatest fear doesn't hold water. If his pf jars contaminated due to contaminated spores/syringe, it does no good to have even ten gallons of contaminated LC on hand. If they contaminated due to his sloppy procedure, he's likely to have used the same sloppy procedure making an LC.

In my posts, I always try to show a way to have the greatest possible success rate, not necessarily the cheapest, easiest or laziest method.
RR




Gotcha. While your technique on making lots of LC will work and is _better_ than just sticking a syringe into an LC, it's just a lot easier for a newb to stick 1cc into a jar to check it out and save the syringe they just spent money on as well as making a quart or whatever of inoculant.

Also, a lot of new growers mess up the water content of their jars and other things involving substrate preparation. I know I did. It's a retarded mistake, but having some LC kept me from losing the strain.

I understand you're going for highest success rate; it's just not feasible for a lot of us here.

Edited by etam (12/07/06 02:32 PM)

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OfflineGrogan
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Re: QUESTION redboy lc [Re: etam]
    #6344807 - 12/07/06 02:38 PM (17 years, 6 months ago)

Not to rob his thread but, roger how do you take a jar of grains quart or pint then make 100 syringes?? Sounds good.


--------------------
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InvisibleBlutjager
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Re: QUESTION redboy lc [Re: RogerRabbit]
    #6344843 - 12/07/06 02:54 PM (17 years, 6 months ago)

Quote:

RogerRabbit said:
Throwing spores into honey or karo has to be the worst possible way to make a liquid culture and I'll be glad when this current fad passes and folks can get back to established techniques.  Just inoculate some grains or brf in the standard way and you'll be miles ahead.

For those who wish to make LC for inoculation, here's a way to get 100 syringes full of inoculant in two weeks: 1) Inoculate a quart jar of rye berries or wbs with your spores.  Upon full colonization, shake the jar to loosen each individual kernel.  Be sure to give it the smell test to make sure it smells like fresh mushrooms. 2) PC a quart of distilled water in a jar with a filter disk or tyvek, etc.  In a glovebox or in front of a flowhood, pour 2/3 of the quart jar of sterilized water into your jar of shaken grains.  3) Shake well and then pour the now mycelium rich water back into whatever was left of your jar of sterilized water. Use the jar lid to hold back the grains themselves from pouring out.  You now have a full quart of highly concentrated mycelium water that can then be used as is, or diluted two or three times again, and used to fill syringes.  Because the grains were shaken first, the mycelium is ripped into shreds and will be sucked through even small needles easily, although larger needles always work better for this.

Remember, mycelium grows poorly in water unless under constant stir which oxygenates it.  With grains, they colonize in two weeks or less because oxygen is throughout the jar in the spaces between the kernels.
RR




This is awesome RR,I plan on giving this a try sometime soon,Never really can have too much lc now can you  :hehehe:  :hehehe:

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Offlineetam
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Re: QUESTION redboy lc [Re: Blutjager]
    #6344851 - 12/07/06 02:58 PM (17 years, 6 months ago)

Quote:

Blutjager said:
Quote:

RogerRabbit said:
Throwing spores into honey or karo has to be the worst possible way to make a liquid culture and I'll be glad when this current fad passes and folks can get back to established techniques.  Just inoculate some grains or brf in the standard way and you'll be miles ahead.

For those who wish to make LC for inoculation, here's a way to get 100 syringes full of inoculant in two weeks: 1) Inoculate a quart jar of rye berries or wbs with your spores.  Upon full colonization, shake the jar to loosen each individual kernel.  Be sure to give it the smell test to make sure it smells like fresh mushrooms. 2) PC a quart of distilled water in a jar with a filter disk or tyvek, etc.  In a glovebox or in front of a flowhood, pour 2/3 of the quart jar of sterilized water into your jar of shaken grains.  3) Shake well and then pour the now mycelium rich water back into whatever was left of your jar of sterilized water. Use the jar lid to hold back the grains themselves from pouring out.  You now have a full quart of highly concentrated mycelium water that can then be used as is, or diluted two or three times again, and used to fill syringes.  Because the grains were shaken first, the mycelium is ripped into shreds and will be sucked through even small needles easily, although larger needles always work better for this.

Remember, mycelium grows poorly in water unless under constant stir which oxygenates it.  With grains, they colonize in two weeks or less because oxygen is throughout the jar in the spaces between the kernels.
RR




This is awesome RR,I plan on giving this a try sometime soon,Never really can have too much lc now can you  :hehehe:  :hehehe:




I skimmed through that the first time.  It really is awesome.  I'll be trying this soon as well, I'll let you know how it turns out!

Thanks again RR

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OfflineRogerRabbitM
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Re: QUESTION redboy lc [Re: etam]
    #6345345 - 12/07/06 06:16 PM (17 years, 6 months ago)

That tek is demonstrated in my upcoming video. Roadkill, who did the pf tek segment used one of those syringes(elm oyster mycelium) to inoculate his pf jars with, which were fully colonized in 8 days at 70F.
RR


--------------------
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semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
Thomas Edison

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Offlinejoker_66599
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Re: QUESTION redboy lc [Re: RogerRabbit]
    #6345396 - 12/07/06 06:28 PM (17 years, 6 months ago)

Quote:

RogerRabbit said:
Throwing spores into honey or karo has to be the worst possible way to make a liquid culture and I'll be glad when this current fad passes and folks can get back to established techniques. Just inoculate some grains or brf in the standard way and you'll be miles ahead.

For those who wish to make LC for inoculation, here's a way to get 100 syringes full of inoculant in two weeks: 1) Inoculate a quart jar of rye berries or wbs with your spores. Upon full colonization, shake the jar to loosen each individual kernel. Be sure to give it the smell test to make sure it smells like fresh mushrooms. 2) PC a quart of distilled water in a jar with a filter disk or tyvek, etc. In a glovebox or in front of a flowhood, pour 2/3 of the quart jar of sterilized water into your jar of shaken grains. 3) Shake well and then pour the now mycelium rich water back into whatever was left of your jar of sterilized water. Use the jar lid to hold back the grains themselves from pouring out. You now have a full quart of highly concentrated mycelium water that can then be used as is, or diluted two or three times again, and used to fill syringes. Because the grains were shaken first, the mycelium is ripped into shreds and will be sucked through even small needles easily, although larger needles always work better for this.

Remember, mycelium grows poorly in water unless under constant stir which oxygenates it. With grains, they colonize in two weeks or less because oxygen is throughout the jar in the spaces between the kernels.
RR




damn got to use this next time

Joker


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?¿
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OfflineAFOD
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Re: QUESTION redboy lc [Re: joker_66599]
    #7409631 - 09/14/07 03:00 PM (16 years, 9 months ago)

I just saw this thread. I have noticed that my jars of colonized rye berries do not simply shake apart. The myc is very dense, even in jars where only 50 or 60 % colonization the myc is so dense that a tool is needed to seperate grains, such as a fork or other instrument. How does this play in? Even as a newb I can see exactly what rr is saying, first you want to be dealing with an isolate and second the finely broken threads of myc will pass through the syringe quite easily. Also, for those who like LC's you could inoculate a jar of Karo or honey lc with one of these syringes and have an pure aggressive strain lc. Why can't we all just get along, there is much to be accomplished by combining these techniques.
As to my particular problem with seperating the grains and breaking the myc, RR amaze us with your almost unequalled wisdom.
Thanks to all, sorry for reviving an old thread.

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