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Toxxy
Random Gary
Registered: 07/07/06
Posts: 36
Last seen: 16 years, 10 months
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Liquid culture air supply
#5903154 - 07/26/06 09:24 PM (17 years, 6 months ago) |
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Okay so my LC (third attempt, this time it's a karo/bottled water mix) finally started going, after about 4 days I had a good cloud about half the size of a ping pong ball. I continued my routine of incubating and shaking, but there's been no further progress. I've got one hole at the top of it that's covered with micropore tape, and that's all it's got for air supply. There's only a half inch of air at the top of the jar (halfpint)
Does it make sense that this would be the cause for my stalled growth, and if so, should I just take the micropore tape off the top and dump some of the solution into the sink, or what? How can I add air to the mix with minimal fear of introducing contams?
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Blutjager
Inhuman


Registered: 06/11/06
Posts: 9,220
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Re: Liquid culture air supply [Re: Toxxy]
#5903189 - 07/26/06 09:32 PM (17 years, 6 months ago) |
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A little LC goes a long way so you probably have quite a useful LC going as it is and just don't realize it,I wouldn't open it and dump any out seeing as you just are asking to let contams in and a bad LC will destroy everything you inoculate with it.Your best bet in my opinion is instead of opening it to let air in you should make a new LC and than use the one you have going to knock that one up and by sucking it up in a syringe you will be allot less likely to expose it to the air.It should take off again once it has more nutes to feed it.I don't think air exchange it the problem anyway,I bet it just used up its fuel and is slowing down.If you feed it it will grow.Hope this helps and Good luck !!
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creamcorn
mad scientist


Registered: 03/13/06
Posts: 2,962
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Re: Liquid culture air supply [Re: Blutjager]
#5903225 - 07/26/06 09:40 PM (17 years, 6 months ago) |
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gas exchange isn't as big a concern with LC as many believe. it looks like a big fluffy cloud of mycelium in there, but fact of the matter is its really not much, and doesnt have very high gas exchange requirements. it looks "fluffed out" because its floating in liquid, but sometime if you want to experiment, pour your LC out through a coffee filter and see how much myc you really have. its deceiving.
that said... unfortunately i've got a less optimistic diagnosis.
when you say shake, are you literally shaking?
if your micropore gets wet with LC solution, its basically a goner. bacteria can grow right through the wet tape. it will then spread extremely quick inside the liquid, and stall your growth.
i use the same lid as you describe. but i fill jars half way to 2/3rds max. agitation is always a very careful swirling motion, to be sure the tape never gets wet.
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liamtheloser
Advanced Idiot

Registered: 06/07/06
Posts: 1,453
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Re: Liquid culture air supply [Re: Blutjager]
#5903232 - 07/26/06 09:41 PM (17 years, 6 months ago) |
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a halfpint will only grow a small cloud like that... the whole thing wont get colonized. i have 12 lc's right now in my fridge, 4 of them are half-pints, when they stopped getting bigger i tossed them in the fridge. And they sound like that look just like yours 
the full pint lc's look like the amount a halfpint SHOULD make if it fully colonized the lc, and i have 1 quart sized that has a mass about the size of 3/4's of a pint jar full... lookin good thouh
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kristen
Burn out..don't fade away

Registered: 04/19/06
Posts: 303
Last seen: 9 years, 25 days
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Re: Liquid culture air supply [Re: liamtheloser]
#5903257 - 07/26/06 09:48 PM (17 years, 6 months ago) |
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Suck up as much of the cloud as you can, it will replenish itself. Shaking them also causes growth slowdown, just gently swirl if you want a huge cloud. Plus 4 days isn't that long, really. Jars in a complete vaccum still work perfectly fine too.
Would you like to smoke some pot, beserker!
Edited by kristen (07/26/06 09:49 PM)
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creamcorn
mad scientist


Registered: 03/13/06
Posts: 2,962
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Re: Liquid culture air supply [Re: kristen]
#5903276 - 07/26/06 09:52 PM (17 years, 6 months ago) |
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yeah thats one point i missed. you said you got good growth at 4 days, then it seemed to stop. well how long has it been since then? if it hasn't gone anywhere in several days and you're certain its stalled, it could be that its just done, or it could be contamed. im a little skeptical that you only got the size of a half ping pong ball and its done though, unless you used a very very small amount of karo.
guess there's only one way to find out and knock up a sample jar...
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Yamidude
Stranger

Registered: 06/15/06
Posts: 957
Last seen: 15 years, 5 months
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Re: Liquid culture air supply [Re: creamcorn]
#5903294 - 07/26/06 09:56 PM (17 years, 6 months ago) |
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my LC is weaksauce right now. I did it over the weekend. Im hoping by this weekend it's much more fluffed up..
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Toxxy
Random Gary
Registered: 07/07/06
Posts: 36
Last seen: 16 years, 10 months
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Re: Liquid culture air supply [Re: kristen]
#5903307 - 07/26/06 09:58 PM (17 years, 6 months ago) |
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This'll probably sound like a dumb question, but how exactly am I supposed to suck up the mycelium in the jar? The needle only gets like a half inch into the jar, the mycelium floats just above the bottom.
Teks I've read just say 'just suck up as much as you can get,' but I don't think my needle will reach.
Edited by Toxxy (07/26/06 09:59 PM)
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creamcorn
mad scientist


Registered: 03/13/06
Posts: 2,962
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Re: Liquid culture air supply [Re: Yamidude]
#5903309 - 07/26/06 09:59 PM (17 years, 6 months ago) |
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only 100ml worth. only swirled it around 2-3 times, left it mostly untouched. nothing in there to agitate (no marble or anything). nothing but a small hole with micropore tape. took about 10 days, maybe slower than some, but i wasn't in a hurry.
one bigass cloud. the secret is in getting the amount of karo right (tho this one is honey, not karo.)
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creamcorn
mad scientist


Registered: 03/13/06
Posts: 2,962
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Re: Liquid culture air supply [Re: Toxxy]
#5903316 - 07/26/06 10:02 PM (17 years, 6 months ago) |
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Quote:
Toxxy said: This'll probably sound like a dumb question, but how exactly am I supposed to suck up the mycelium in the jar? The needle only gets like a half inch into the jar, the mycelium floats just above the bottom.
Teks I've read just say 'just suck up as much as you can get,' but I don't think my needle will reach.
tilt the jar on an angle. 
careful that it doesnt spill out the hole. suck up a little, squirt back in, and it breaks it up into lots of little bits, then suck up again slowly and you'll capture plenty of bits.
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Yamidude
Stranger

Registered: 06/15/06
Posts: 957
Last seen: 15 years, 5 months
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Re: Liquid culture air supply [Re: creamcorn]
#5903317 - 07/26/06 10:02 PM (17 years, 6 months ago) |
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how far along was that pic taken CC?
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creamcorn
mad scientist


Registered: 03/13/06
Posts: 2,962
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Re: Liquid culture air supply [Re: Yamidude]
#5903323 - 07/26/06 10:03 PM (17 years, 6 months ago) |
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Quote:
Yamidude said: how far along was that pic taken CC?
10 days. i edited my post to include that probably as you were asking this.
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Yamidude
Stranger

Registered: 06/15/06
Posts: 957
Last seen: 15 years, 5 months
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Re: Liquid culture air supply [Re: creamcorn]
#5903336 - 07/26/06 10:06 PM (17 years, 6 months ago) |
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Quote:
creamcorn said:
Quote:
Yamidude said: how far along was that pic taken CC?
10 days. i edited my post to include that probably as you were asking this.
good to know, mine are only a few days along. i have 2 TX and one B+. i need some new shit. I have like 70billion trays going of B+.
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Tippinthru
contented

Registered: 04/07/05
Posts: 1,131
Loc: "The Garden"...
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Re: Liquid culture air supply [Re: Toxxy]
#5903690 - 07/26/06 11:27 PM (17 years, 6 months ago) |
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Quote:
Toxxy said: This'll probably sound like a dumb question, but how exactly am I supposed to suck up the mycelium in the jar? The needle only gets like a half inch into the jar, the mycelium floats just above the bottom.
Teks I've read just say 'just suck up as much as you can get,' but I don't think my needle will reach.
Use a BIGGER LONGER needle 
 (joke)
  1000 ml
-------------------- Perfection is attained by slow degrees; it requires the hand of time... [
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Omnicracker
Crusted Trolltivator

Registered: 09/18/05
Posts: 1,421
Loc: Wal-Merica
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Re: Liquid culture air supply [Re: kristen]
#5903787 - 07/26/06 11:43 PM (17 years, 6 months ago) |
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Quote:
kristen said: Jars in a complete vaccum still work perfectly fine too.
have fun aspirating the LC without a filter port. that LC has to displace with something... how about some contams!
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Yamidude
Stranger

Registered: 06/15/06
Posts: 957
Last seen: 15 years, 5 months
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Re: Liquid culture air supply [Re: Tippinthru]
#5903799 - 07/26/06 11:45 PM (17 years, 6 months ago) |
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Quote:
Tippinthru said:
Quote:
Toxxy said: This'll probably sound like a dumb question, but how exactly am I supposed to suck up the mycelium in the jar? The needle only gets like a half inch into the jar, the mycelium floats just above the bottom.
Teks I've read just say 'just suck up as much as you can get,' but I don't think my needle will reach.
Use a BIGGER LONGER needle 
1000 ml
do you use a stirrer?
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Tippinthru
contented

Registered: 04/07/05
Posts: 1,131
Loc: "The Garden"...
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Re: Liquid culture air supply [Re: Omnicracker]
#5903820 - 07/26/06 11:49 PM (17 years, 6 months ago) |
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Quote:
Omnicracker said:
Quote:
kristen said: Jars in a complete vaccum still work perfectly fine too.
have fun aspirating the LC without a filter port. that LC has to displace with something... how about some contams!
TRUE THAT.
Or, make your LC in 3000 ml IV bags. 
-------------------- Perfection is attained by slow degrees; it requires the hand of time... [
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Omnicracker
Crusted Trolltivator

Registered: 09/18/05
Posts: 1,421
Loc: Wal-Merica
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Re: Liquid culture air supply [Re: Tippinthru]
#5903826 - 07/26/06 11:51 PM (17 years, 6 months ago) |
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thats sexy
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Tippinthru
contented

Registered: 04/07/05
Posts: 1,131
Loc: "The Garden"...
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Re: Liquid culture air supply [Re: Yamidude]
#5903957 - 07/27/06 12:17 AM (17 years, 6 months ago) |
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>>>>>>>do you use a stirrer?
Yes 
Galvanized nail as a stir bar 
-------------------- Perfection is attained by slow degrees; it requires the hand of time... [
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kristen
Burn out..don't fade away

Registered: 04/19/06
Posts: 303
Last seen: 9 years, 25 days
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Re: Liquid culture air supply [Re: Tippinthru]
#5904395 - 07/27/06 04:42 AM (17 years, 6 months ago) |
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You can pc an empty syringe and inject sterile air, use a polyfil needle to de-pressurize, or flame air to suck in the syringe before pulling out the culture.
But anyway, the point was LC's don't need massive air exchange and will still work.
And I haven't had contams in a long time, thanks.
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