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 HighHat
USDA Certified,Grade A 3===D
Registered: 01/24/08
Posts: 269
Loc: I TiVo my eBay. Constantl...
Last seen: 3 days, 16 hours
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 GM Giving Me No Breaks... Ideas?
#8533346 - 06/17/08 11:24 AM (2 months, 20 days ago) |
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Ok, so Ive been attempting a grow for a few months now, but every time I get to incubating my rye jars, myc starts to grow nicely then all of the sudden a giant patch of green starts spreading from the center of the myc outward.
Here's what I've been doing:
I made a (roughly) 3% honey/distilled water LC, let it sit for 12 hours (to germinate endospores) using rolled pieces of polyfill batting pulled through a small hole for a gas exchange filter, PC'd it for 35 min, then let it cool in the PC. After the pressure dropped in the PC, I placed a piece of polyfill batting soaked in alcohol over the regulator vent to sterilize the incoming air.
After cooling, I placed the LC, spore print (never opened, still in vendor bag), and boiled tweezers in a clear rubber maid glovebox with arm holes melted into the side. I then sprayed a generous amount of Oust Air+Surface cleaner into the box, put the lid on, then covered the armholes with saran wrap. The glovebox was then left to sit for about 10min.
Using home made long rubber gloves (medium length cleaning gloves with the hand cut off and a regular latex glove on my hands with the cuff sealed with duct tape) which I then rubbed down with alcohol before inserting them into the glovebox. I scraped some spores out of the print bag and dropped them into the LC. I re-sealed the LC, then put it into a TIT incubator set for 84F.
After awhile, myc formed in the LC, which I drew out under the same sterile conditions with a new (Vet sterile) 16G 12cc syringe through a silicone injection port I made with high temperature silicone over a hole I punched in the LC lid (before PCing).
I then prepared rye jars (250ml dry grain : 180ml distilled water) with home-made silicone injection ports which were PC'd for 90min. The same sterile technique was used for the rye jars. Each time I took the cap off the needle, I flamed it until it was glowing red, then wiped it with a piece of paper towel soaked with alcohol right before injecting the rye jars.
The rye jars were then incubated at 86F until contaminated.
I have done everything I know of (aside from making a flow hood) to figure out where the contaminants are getting in. My apartment is new, the carpet was just cleaned, I keep everything clean.
I decided to make a spore syringe and start back from a multispore inoculation of an LC, just in case my LC is contaminated, but I cant even get any new spores to germinate in any new LC's.
I PC'd a syringe (same size), tweezers, and a 60ml beaker, put everything in the glovebox, sterilized the inside, scraped some spores into the beaker, then squirted sterilized distilled water from the syringe into the beaker and drew it back out (flaming the needle and rubbing it with alcohol before and after removing the cap).
So where I'm at is I have a spore syringe that I'm beginning to think may be contaminated because no spores will germinate (even after incubating the LC for 2 and a half weeks) and a myc syringe that WILL grow more myc in a new LC, but every time I try to use the myc to inoculate rye jars, even through injection ports, contaminates with Green mold.
Is there something that I'm missing?
If my LC syringe is contaminated, along with my spores, is there any way to isolate some clean myc? Agar work? Cardboard isolation?
Thanks.
EDIT: I also have heard that the water in the bottom of the TIT can be responsible for contams, so I cleaned out the tubs, put fresh water in the bottom, and added a good bit of H2O2 to the bottom to prevent its contamination. I did this about 5 weeks ago.
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Have you ever felt like you were wearing a hat, but you weren't?
"Those who would give up Essential Liberty to purchase a little Temporary Safety, deserve neither Liberty nor Safety" -Letter from the Pennsylvania Assembly, November 11, 1755
This profile is strictly for role-playing. Any alleged association with illegal activities is purely fictional.
Edited by HighHat (06/17/08 11:27 AM)
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 RogerRabbit
Bans forPleasure
Registered: 03/26/03
Posts: 16,870
Loc: Pacific Northwest
Last seen: 24 seconds
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 Re: GM Giving Me No Breaks... Ideas? [Re: HighHat]
#8535490 - 06/17/08 10:30 PM (2 months, 19 days ago) |
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I strongly recommend against making LC from spores. While it seems to be all the rage, you don't even know if you have contamination until you've wasted all the time to grow the LC, and then you have to use it and wait to see if it's any good.
Simply use your spores straight to grains and you'll know in a week or two if it's contaminated. Even better is to swipe the spores onto agar in a petri dish, and then you can clean up and isolate healthy mycelium away from the contamination.
In addition, don't incubate. Heat, especially above 81 -83F, is going to slow down your mycelium and encourage molds.
RR
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www.mushroomvideos.com
"I wouldn't want to belong to any club that would have me as a member".
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HighHat
USDA Certified,Grade A 3===D
Registered: 01/24/08
Posts: 269
Loc: I TiVo my eBay. Constantl...
Last seen: 3 days, 16 hours
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Re: GM Giving Me No Breaks... Ideas? [Re: RogerRabbit]
#8536982 - 06/18/08 12:05 PM (2 months, 18 days ago) |
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When you say not to incubate, does that go for sub jars too, or just LC's? I thought 86F was the optimal temperature for mycellial growth, but I was also aware that the higher temperature encourages contaminates too. Are you suggesting that I ditch the TIT and just put my jars in a rubber maid to colonize at room temperature? (about 76F) I did notice some crazy rhizomorphs once a while ago I left a slightly contaminated jar out in the room overnight, I thought that it was just a coincidence.
I'll be getting the materials for agar work soon, hopefully I'll be able to sort this mess out without having to buy new spores. One bad possibility that I am considering is I may have opened the spore print while there was still Oust floating around in the glovebox... Might that have killed the print?
Either way, Ill try going straight to grain this week and see what happens.
Much appreciated RR.
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Have you ever felt like you were wearing a hat, but you weren't?
"Those who would give up Essential Liberty to purchase a little Temporary Safety, deserve neither Liberty nor Safety" -Letter from the Pennsylvania Assembly, November 11, 1755
This profile is strictly for role-playing. Any alleged association with illegal activities is purely fictional.
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endepth
The world remains ever the same.
Registered: 12/25/07
Posts: 630
Loc: Cloud 9
Last seen: 1 day, 21 hours
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Re: GM Giving Me No Breaks... Ideas? [Re: HighHat]
#8537373 - 06/18/08 02:58 PM (2 months, 18 days ago) |
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Old info. Just ditch the incubator all together. Room temp is fine.
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Being normal is nothing to aspire to. It is something to get away from.
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The shroomy 1
Let me be YOUR"guide"
Registered: 03/27/07
Posts: 1,876
Loc: Texas
Last seen: 14 hours, 10 minutes
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Re: GM Giving Me No Breaks... Ideas? [Re: HighHat]
#8538182 - 06/18/08 06:58 PM (2 months, 18 days ago) |
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I have a few F+ prints available if you need one. No trade, no worries. If you are interested, PM me.
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