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 legallyhomeless
mooch
Registered: 07/01/05
Posts: 4,051
Loc: EZRA for the Refuge
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 LC always contaming my jars 
#7231149 - 07/27/07 04:19 PM (17 years, 2 months ago) |
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I made some nice LC (karo + distilled)  I preped some WBS, rinsed, soaked 24, rinsed, drained 6. But every time I use my LC to innoc a jar it contams within 2 days or less. Last time it was cobweb and this time its the green. IDK wtf to do. I asume that the LC is making the WBS too wet or something. Any words?
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 The shroomy 1
Luminous beings surround me
Registered: 03/27/07
Posts: 5,543
Loc: The Aether
Last seen: 4 months, 1 day
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Quote:
legallyhomeless said:
I preped some WBS, rinsed, soaked 24, rinsed, drained 6.
Did I miss the part where you say, "I then preasure cooked the wbs for x amount of time at 15psi."?
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AMU Q&A thread.
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Brainiac
Rogue Scientist
Registered: 04/29/06
Posts: 13,259
Loc: 與您的女朋 
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Maybe the lc is contemned.
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 Fair is Fair 
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simplemachine
Manfly
Registered: 09/14/03
Posts: 1,981
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That could be the case, but I think its more likely that there are contams swimming in the lc that you cannot see. I have had some pretty lousy luck with LCs, so I prefer to use agar and G2G for cloning, isolating, and propagating.
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The shroomy 1
Luminous beings surround me
Registered: 03/27/07
Posts: 5,543
Loc: The Aether
Last seen: 4 months, 1 day
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Re: LC always contaming my jars [Re: Brainiac]
#7231262 - 07/27/07 04:46 PM (17 years, 2 months ago) |
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Quote:
Brainiac said:
Maybe the lc is contemned.
The liquid culture looks fine. I don't see any contamination in the jar! The soaking of the wbs is to help germinate the endospores that are present in the wbs. If you don't pc the wbs your left with lots of nasties just waiting to colonize. YOU NEED TO PRESSURE COOK the wbs and finish what you started.
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AMU Q&A thread.
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ThePyschonaut52
Stranger
Registered: 04/24/07
Posts: 982
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Re: LC always contaming my jars [Re: The shroomy 1]
#7231305 - 07/27/07 04:57 PM (17 years, 2 months ago) |
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i never pc! i after making grain/jars, i always go take a shit, dont wash my hands, and finger bang my cakes. i then sneeze on the grain/substrate, then leave everything outside(with the lids off, of course). THEN i inject spores!!
just kidding...
but seriously, if you dont follow sterile procedures, like flaming and swabbing your needle with alcohol before/after using your LC syringe, then your likely gonna get contams...
good luck!
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"In god we trust..."
-I guess we're screwed.
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The shroomy 1
Luminous beings surround me
Registered: 03/27/07
Posts: 5,543
Loc: The Aether
Last seen: 4 months, 1 day
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Quote:
ThePyschonaut52 said:
i never pc! i after making grain/jars, i always go take a shit, dont wash my hands, and finger bang my cakes. i then sneeze on the grain/substrate, then leave everything outside(with the lids off, of course). THEN i inject spores!!
just kidding...
but seriously, if you dont follow sterile procedures, like flaming and swabbing your needle with alcohol before/after using your LC syringe, then your likely gonna get contams...
good luck!
you actually had me going there at first! LOL!! I was thinking to myself. "Oh my God! How does he not get contaminations?"
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AMU Q&A thread.
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ThePyschonaut52
Stranger
Registered: 04/24/07
Posts: 982
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Re: LC always contaming my jars [Re: The shroomy 1]
#7231565 - 07/27/07 06:33 PM (17 years, 2 months ago) |
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 i felt like being goofy, lol
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"In god we trust..."
-I guess we're screwed.
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xaxphaanes
Mycologist
Registered: 08/08/05
Posts: 2,988
Last seen: 2 years, 2 months
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Re: LC always contaming my jars [Re: The shroomy 1]
#7231571 - 07/27/07 06:36 PM (17 years, 2 months ago) |
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Quote:
The shroomy 1 said:
Quote:
Brainiac said:
Maybe the lc is contemned.
The liquid culture looks fine. I don't see any contamination in the jar! The soaking of the wbs is to help germinate the endospores that are present in the wbs. If you don't pc the wbs your left with lots of nasties just waiting to colonize. YOU NEED TO PRESSURE COOK the wbs and finish what you started.
It's almost impossible to see a contamination in a lc.
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"Anything i say is fictional"
what you should look for in manure
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legallyhomeless
mooch
Registered: 07/01/05
Posts: 4,051
Loc: EZRA for the Refuge
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Sorry I was at work and in a hurry. OF COURSE I PC 15 psi for 2 hrs, youde have to be a moron not to  I thought the LC looked fine too, Ive had some bad luck too but this was a nice batch
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xaxphaanes
Mycologist
Registered: 08/08/05
Posts: 2,988
Last seen: 2 years, 2 months
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1 hour that's still a long pc time for an lc.I do mine at 15psi for 30 mins
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"Anything i say is fictional"
what you should look for in manure
Edited by xaxphaanes (07/27/07 08:25 PM)
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legallyhomeless
mooch
Registered: 07/01/05
Posts: 4,051
Loc: EZRA for the Refuge
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Re: LC always contaming my jars [Re: The shroomy 1]
#7231821 - 07/27/07 08:05 PM (17 years, 2 months ago) |
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Quote:
The shroomy 1 said:
Quote:
legallyhomeless said:
I preped some WBS, rinsed, soaked 24, rinsed, drained 6.
Did I miss the part where you say, "I then preasure cooked the wbs for x amount of time at 15psi."?
I PC the WBS for 2 hours. The LC for about 1 hr
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The shroomy 1
Luminous beings surround me
Registered: 03/27/07
Posts: 5,543
Loc: The Aether
Last seen: 4 months, 1 day
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Seems like the syringe is your culprit, along with maybe the sterile technique during inoculation.
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AMU Q&A thread.
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RogerRabbit
Bans for Pleasure
Registered: 03/26/03
Posts: 42,214
Loc: Seattle
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Re: LC always contaming my jars [Re: The shroomy 1]
#7232918 - 07/28/07 06:19 AM (17 years, 2 months ago) |
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Use agar to verify your spores, not LC. LC should not be made except with a proved, isolated fruiting strain. You'll rarely see contaminants in LC because molds generally won't sporulate in water.
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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legallyhomeless
mooch
Registered: 07/01/05
Posts: 4,051
Loc: EZRA for the Refuge
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Re: LC always contaming my jars [Re: RogerRabbit]
#7233659 - 07/28/07 01:04 PM (17 years, 2 months ago) |
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But how could I be getting a different contam each time I use it? I asure you my syringes are steril along with my innocing techniques. I like to keep shit very very clean. Flame needle, alcohol needle, alcohol injection site, put needle through alcohol wet injection site while I have an alcohol soaked paper towel around the needle. Then pull the needle out and leave papertowel over the hole until i can cover with micropore. Never have had a contam using this method unless of course Im talking about LC them it contams.
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ThePyschonaut52
Stranger
Registered: 04/24/07
Posts: 982
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do you have self-healing injection ports? if not, you can make them; just go to the hardware store and buy some heat-resistant silicon(only a couple bucks for a tube)
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"In god we trust..."
-I guess we're screwed.
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aspore
myconaut
Registered: 07/10/07
Posts: 663
Last seen: 12 years, 1 month
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did you say you PC your LC for 1 hour? That's cool, I am afraid of overdoing mine.
Sounds like your LC is definely bad, I once tried to use baby bottles for some of my first LC"s and those bitches got so infected with shit, but they looked perfectly fine.
I suggest making a fresh master LC, then by taking just one syringe of myc out, making a small army of clone LCs. Just so you don't have to keep poking your master culture, possibly introducing contams.
How was your LC prepared exactly, and how many times have you stuck a needle in it?
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c l i c k m e
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legallyhomeless
mooch
Registered: 07/01/05
Posts: 4,051
Loc: EZRA for the Refuge
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Re: LC always contaming my jars [Re: aspore]
#7234671 - 07/28/07 06:55 PM (17 years, 2 months ago) |
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Ive PCed LC for 2 hrs before and its been fine. As everyone can see by my first post the LC is fine. My LC was made by pouring light karo into distilled or maybe it was spring water. IDK how much, just enough. Maybe 8TBS to 900 ML water. I have a brand new LC that hasnt been poked yet. But again... the LC is fine. The other LC I was using was only poked twice and the first time the jars I innoced with got cobweb and now this time (trich?) that green shit
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Brainiac
Rogue Scientist
Registered: 04/29/06
Posts: 13,259
Loc: 與您的女朋
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Re: LC always contaming my jars [Re: RogerRabbit]
#7234925 - 07/28/07 08:42 PM (17 years, 2 months ago) |
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Quote:
RogerRabbit said:
Use agar to verify your spores, not LC. LC should not be made except with a proved, isolated fruiting strain. You'll rarely see contaminants in LC because molds generally won't sporulate in water.
RR
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Fair is Fair
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aspore
myconaut
Registered: 07/10/07
Posts: 663
Last seen: 12 years, 1 month
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Re: LC always contaming my jars [Re: Brainiac]
#7234950 - 07/28/07 08:52 PM (17 years, 2 months ago) |
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whank kind of lids are you using for your jars? You might have a leak.
bottled water is a must, try getting a cheap gallon
Perhaps your LC was infected via spore injection, perhaps a bad spore syringe.
Try this though, PC up some jars, but don't shoot them up for a week, see if the nasties grow, if not, you know it's your LC. Or just shoot up one jar.
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c l i c k m e
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