|
Some of these posts are very old and might contain outdated information. You may wish to search for newer posts instead.
|
RandomFX
protege



Registered: 12/02/13
Posts: 1,015
Loc: North-East, USA
Last seen: 5 years, 1 month
|
Re: Liquid Culture De-Mystified: Ghia Style [Re: ghiajake]
#19599907 - 02/21/14 02:29 PM (9 years, 11 months ago) |
|
|
Quote:
ghiajake said: I did make a typo, it was 2 out of 6, with 4 still good. To be fair, I did noc all 6 with grains from jars I already G2G'd other grains with, so that's most likely the cause. If I had used agar I'd be throwing away money (dishes) right now instead of just dumping them and trying again.
Fair enough, but that is still 33% but then again it is water and myc...not like a major expense. As far as the dishes. use glass petri's. I do, and do not throw any of them away, and I have two sleeves of disposables that I haven't opened since. cynmar.com sells glass petri's for .80 a piece in packs of 10. for 15mm x 75mm I believe.
|
ghiajake
Myco-Viking


Registered: 01/10/13
Posts: 3,846
Loc: Indiana
Last seen: 14 hours, 6 minutes
|
Re: Liquid Culture De-Mystified: Ghia Style [Re: RandomFX]
#19600109 - 02/21/14 03:18 PM (9 years, 11 months ago) |
|
|
Glass petris are awesome, but I can't do a liquid extraction like with the jars. I will probably pick some up eventually, just because they're cool. I picked up 5x 2ml glass syringes for that very same reason. Gonna order a 100mL one eventually too.
|
kotter


Registered: 01/15/11
Posts: 210
Last seen: 4 years, 7 months
|
Re: Liquid Culture De-Mystified: Ghia Style (moved) [Re: ghiajake]
#19664651 - 03/07/14 06:40 PM (9 years, 10 months ago) |
|
|
Maybe LC does have limited usefulness but it enabled this clone to enter my life:
On grain: two jars of erinaceus; a jar of corolloides is on the right.

erinaceus

No idea why I could not get the above two not to rotate. I unchecked the pertinent box to no avail.
This is my very first lion's mane fruit from a clone I made!
Several more bags are close behind (just as I'm leaving town for a week)


The original fruiting body it came from:
Edited by kotter (03/07/14 06:41 PM)
|
blindingleaf
blue collar underworld



Registered: 07/19/13
Posts: 22,008
Loc: sub-surface unseen
|
Re: Liquid Culture De-Mystified: Ghia Style (moved) [Re: kotter]
#19664661 - 03/07/14 06:42 PM (9 years, 10 months ago) |
|
|
nice pics kotter!
-------------------- A few thoughts on cultivation MICROBIAL HUSBANDRY!!!! The whole is greater than the sum of its parts
|
kotter


Registered: 01/15/11
Posts: 210
Last seen: 4 years, 7 months
|
Re: Liquid Culture De-Mystified: Ghia Style (moved) [Re: blindingleaf]
#19664829 - 03/07/14 07:34 PM (9 years, 10 months ago) |
|
|
Thanks! I'm still pretty low on the learning curve but have sure become obsessed with Hericiums. I've halfway filled a room with bags on racks. I'll post more images when I get back home.
|
invitro


Registered: 05/03/13
Posts: 2,529
Last seen: 1 month, 19 days
|
Re: Liquid Culture De-Mystified: Ghia Style [Re: x7x_x7x]
#19664855 - 03/07/14 07:39 PM (9 years, 10 months ago) |
|
|
Quote:
x7x_x7x said: read SuctoSpore® Pictorial Tek
I read this tek, but I don't really see the advantage of using a 3 way stop cock. I've never seen one up-close but my main concern is how do you sterilize the part of the stop cock that receives the needle? If your familiar with Tl's no tilt lc tek, this seems to offer the same functionality and the rtv blob is easy to sterilize with alcohol.
http://www.shroomery.org/forums/showflat.php/Number/14186384
|
ghiajake
Myco-Viking


Registered: 01/10/13
Posts: 3,846
Loc: Indiana
Last seen: 14 hours, 6 minutes
|
Re: Liquid Culture De-Mystified: Ghia Style [Re: invitro]
#19664886 - 03/07/14 07:47 PM (9 years, 10 months ago) |
|
|
Nice looking culture and fruits kotter!
invitro: I briefly debated using female luer-lock tips for my LC lids, but not with a check valve like that. I was going to use caps for them instead. Didn't use them because I already had the parts for the no-tilts I made.
|
invitro


Registered: 05/03/13
Posts: 2,529
Last seen: 1 month, 19 days
|
Re: Liquid Culture De-Mystified: Ghia Style [Re: ghiajake]
#19664956 - 03/07/14 08:13 PM (9 years, 10 months ago) |
|
|
What I really like about the rtv blobs is that you can wipe alcohol on them wet, with a small square of sponge, then ring out the same sponge and kind of dab it 'dry' Leaving what is probably a very thin film of alcohol which probably evaporates nearly instantly. The evaporation of alcohol from a contam spore is actually what kills it, bursting it from the inside (I read that somewhere). A 3 way port or a luer lock would just kind of stay wet. I'm not sure if having it stay wet is actually a disadvantage but it seems like it might be. Someone who is more up on science would have to chime in there.
|
bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,889
Loc: Milky way
|
Re: Liquid Culture De-Mystified: Ghia Style [Re: invitro]
#19665115 - 03/07/14 09:04 PM (9 years, 10 months ago) |
|
|
Silicone injection ports are always a risk. The best you can do is wipe them with alcohol which is only a sanitizer not a sterilizer so you always have a small chance of pushing contams through no matter what. Same reason why no one wipes with alcohol after flaming it's pointless. Alcohol than flame, You can't flame a silicone injection port so you're always trying your luck and every single time you use a LC you have to test it.
|
Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,808
Loc: Canada
|
Re: Liquid Culture De-Mystified: Ghia Style [Re: bodhisatta]
#19665140 - 03/07/14 09:12 PM (9 years, 10 months ago) |
|
|
That's why I use as large a syringe as possible and crack the lid when inoculating the cut down those vectors. Though even a 60 cc syringe may test clean on the first couple jars, but after doing 10-12 more it might have picked up a contam for the last one or two. That's why you must treat LC like a grenade. It has great power, but can go off in yer face if your not careful.
|
YOmamaPr0
KnottyBetch



Registered: 12/19/13
Posts: 780
Loc: New Mexico, US
Last seen: 1 year, 8 months
|
Re: Liquid Culture De-Mystified: Ghia Style [Re: Pastywhyte]
#19665185 - 03/07/14 09:26 PM (9 years, 10 months ago) |
|
|
Like a grenade. Metaphorically beautiful. *technically a simile but it didn't sound as good.
|
Forrester
aspiring sociopath


Registered: 02/05/13
Posts: 9,351
Loc: Northeast USA
Last seen: 24 days, 2 hours
|
Re: Liquid Culture De-Mystified: Ghia Style [Re: bodhisatta]
#19665199 - 03/07/14 09:31 PM (9 years, 10 months ago) |
|
|
Quote:
bodhisatta said: Silicone injection ports are always a risk. The best you can do is wipe them with alcohol which is only a sanitizer not a sterilizer so you always have a small chance of pushing contams through no matter what. Same reason why no one wipes with alcohol after flaming it's pointless. Alcohol than flame, You can't flame a silicone injection port so you're always trying your luck and every single time you use a LC you have to test it.
Very good point - unless, you just don't remove the foil after PC, until cooled and ready to inject. Foil off, BAM! inject. Pretty sterile-like
-------------------- Repugnant is a creature who would squander the ability to lift an eye to heaven, conscious of his fleeting time here. ------------------- Have some medicinal mushrooms and want to get the most out of them? Try this double extraction method.
|
Mrcloudy
Stranger than you.



Registered: 10/01/13
Posts: 2,889
Loc: Northeast US
Last seen: 3 months, 18 days
|
Re: Liquid Culture De-Mystified: Ghia Style [Re: bodhisatta]
#19665227 - 03/07/14 09:36 PM (9 years, 10 months ago) |
|
|
So I am new at working with Hericium, not to detract from the conversation about Ghias liquid culture. But is that whispy branching common in hereciums? Several months ago I found a dried sad looking mushroom fruiting from pine and identified it as Hericium americanum. Since this was the only one I could find that had been collected from conifers before. By some bizzare miracle, perhaps due to the mother mushroom being immature I was able to clone and clean up a culture without the nightmare that I have read about with cloning lions manes. I had zero bacterial issues, but a few fungal contams.
I have been apprehensive about growing it out because I have suspected it was a contaminant. The culture has been sitting in my fridge this whole time because I have a terrible habit of doing that. But I guess what I want to know is does this look like Herecium mycelium? It has a similar whispy branchlike growth thing going on. I have been meaning to grow it out for testing but havent gotten around to it yet and didn't want to waste time trying to grow something that wasn't what I was after.

Again sorry, I don't mean to detract from this thread, I have just been wondering about my culture for a while now and that jar pic there reminded me I still have this thing hanging out in there.
--------------------
10 different Ganoderma species from across the USA AMU MrCloudys guide to North American GanodermaUpdated A rough guide to North American Ganoderma species, with an emphasis on the laccate species.
|
ghiajake
Myco-Viking


Registered: 01/10/13
Posts: 3,846
Loc: Indiana
Last seen: 14 hours, 6 minutes
|
Re: Liquid Culture De-Mystified: Ghia Style [Re: Mrcloudy]
#19665248 - 03/07/14 09:42 PM (9 years, 10 months ago) |
|
|
Detract away my friend.
|
invitro


Registered: 05/03/13
Posts: 2,529
Last seen: 1 month, 19 days
|
Re: Liquid Culture De-Mystified: Ghia Style [Re: Forrester]
#19665304 - 03/07/14 09:55 PM (9 years, 10 months ago) |
|
|
Quote:
Forrester said:
Quote:
bodhisatta said: Silicone injection ports are always a risk. The best you can do is wipe them with alcohol which is only a sanitizer not a sterilizer so you always have a small chance of pushing contams through no matter what. Same reason why no one wipes with alcohol after flaming it's pointless. Alcohol than flame, You can't flame a silicone injection port so you're always trying your luck and every single time you use a LC you have to test it.
Very good point - unless, you just don't remove the foil after PC, until cooled and ready to inject. Foil off, BAM! inject. Pretty sterile-like 
Yea that's a good idea, plus if your needle goes into the ship when the needle was red-hot 2 seconds ago, anything that might get "pushed through" is probably going to get burned to death on the way in. You wouldn't think the silicone could handle that kind of heat but the red rtv silicone can take it. You might singe it a little though.
Edited by invitro (03/07/14 09:56 PM)
|
bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,889
Loc: Milky way
|
Re: Liquid Culture De-Mystified: Ghia Style [Re: invitro]
#19665336 - 03/07/14 10:01 PM (9 years, 10 months ago) |
|
|
even regular silicone will handle it, Silicone caulking is all 100% silicone and can easily take the red hot needle, the RTV just means it cures a bit differently (room temperature vulcanizing IE could cure underwater and without fresh air) you can cure it in the PC as soon as it forms a skin instead of waiting 24+ hours. The needle cools quite a bit very quickly and it has a low thermal mass, temperature doesn't kill bacteria temperature+time kills bacteria so it's very easy for the bacteria to survive the hot needle poke if the port is dirty, or the alcohol failed to kill something. It's nothing to get crazy worried about but if things go wrong, it's one of the possibilities. This is why most of us don't bother with LC, LC is good for vendors to sell edible/medicinal tried and tested monoculture rather than having you grow from spores and get average genetics, it doesn't mean it's a great way to inoculate and store inoculant, especially for actives like cubes.
Edited by Trusted cuItivator (03/07/14 10:03 PM)
|
invitro


Registered: 05/03/13
Posts: 2,529
Last seen: 1 month, 19 days
|
Re: Liquid Culture De-Mystified: Ghia Style [Re: bodhisatta]
#19665371 - 03/07/14 10:12 PM (9 years, 10 months ago) |
|
|
Do you have a reference that could expand on this idea of heat + time kills bacteria. I can understand maybe if the temperature increase was gradual you might say that, like starting up a pot of boiling water or a pressure cooker. But lets say there is some bacteria on the ship and the still very hot needle hits it, that kind of instant temperature change would undoubtedly decrease the amount of time needed to kill the bacteria dramatically don't you think? Kind of an analogy would be glass jars, you can heat them up in a pc over 30 minutes and they won't break, but try applying rapid heat, like a flame, to the jar, it'll shatter.
|
bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,889
Loc: Milky way
|
Re: Liquid Culture De-Mystified: Ghia Style [Re: invitro]
#19665429 - 03/07/14 10:26 PM (9 years, 10 months ago) |
|
|
needles are hollow bacteria are microscopic the area around the needle will have very slow heat transfer through the silicone and then the jabbing could dislodge or rub them against the inside of the needle which has cooled down who knows but peoples LC contaminates even when made from agar wedges all the time having been clean for a few weeks.
direct flame will oxidase/incinerate anything the flame is nearly 2000F from butane if I remember right but you can see how quickly your needle ceases to stay red hot, it's probably a long shot but I don't discount it considering the fate of most LC after repeated stabbings.
|
invitro


Registered: 05/03/13
Posts: 2,529
Last seen: 1 month, 19 days
|
Re: Liquid Culture De-Mystified: Ghia Style [Re: bodhisatta]
#19665446 - 03/07/14 10:32 PM (9 years, 10 months ago) |
|
|
Even though the needle is not red anymore when it hits the port, it's still plenty hot. When I accidentally bump the plunger on the syringe and a drop comes into the needle, the water vaporizes and steam shoots out of the needle tip.
Innoculating a lc with an agar wedge is not smart, I will agree with that. Doing it the way I said seems to be pretty effective IME.
|
bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,889
Loc: Milky way
|
Re: Liquid Culture De-Mystified: Ghia Style [Re: invitro]
#19665459 - 03/07/14 10:34 PM (9 years, 10 months ago) |
|
|
Quote:
invitro said: Even though the needle is not red anymore when it hits the port, it's still plenty hot. When I accidentally bump the plunger on the syringe and a drop comes into the needle, the water vaporizes and steam shoots out of the needle tip.
Innoculating a lc with an agar wedge is not smart, I will agree with that. Doing it the way I said seems to be pretty effective IME.
Inoculating a lc with an agar wedge is not smart*
Agar wedge is the best way, but even then they can still contam and it's likely from the needle poking. Also I suspect a lot of people inadvertently cool their needle by plunger pressing as they stick the injection port
|
|