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globejs
lollygagger



Registered: 08/18/07
Posts: 414
Last seen: 3 months, 21 days
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first g2g transfer success?
#22505499 - 11/10/15 02:52 PM (8 years, 2 months ago) |
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Hey anyone, I was wondering how long it would take to know if I successfully did a g2g transfer. That is to say how many days after would contamination show up. I believe I did them on the 5th and here are some pics, I transferred a fully colonized multispore quart rye berry jar to three corn.
this was done infront of my flow hood in a 5x7 area i plasticed off with 6 mil plastic and taped the seams, i let it run for over an hour and lysoled (with the kind made for air/surface sanitizing) the front of the filter and the walls and air a ton and once again after i entered the room by lifting the plastic sheeting and going under. i alcohol soaked everything from my hands/ jars, hair, etc. and sat in the nauseating alcohol fumes for 20 minutes before beginning the process.
One jar was not shaken after a one inch layer of rye was added to the top in hopes that if contam was introduced, the mycelium would overtake it by growing over it downward rather than shaken into the jar anywhere where the mycelium would not be near it to overtake it. Is that bad logic?
they have been sitting in my tub in tub incubator at about 84 degrees. theres no strong growth, very fuzzy, but its trying!
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NEWT TESTED, NEWT APPROVED.
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spacechildo
proletarians rise up


Registered: 01/24/13
Posts: 19,243
Loc: Babylon
Last seen: 6 years, 4 months
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Re: first g2g transfer success? [Re: globejs]
#22505666 - 11/10/15 03:34 PM (8 years, 2 months ago) |
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most of those look OK so far, hope you got a proper filter underneath the cloth thing.
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globejs
lollygagger



Registered: 08/18/07
Posts: 414
Last seen: 3 months, 21 days
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Re: first g2g transfer success? [Re: spacechildo]
#22505736 - 11/10/15 03:54 PM (8 years, 2 months ago) |
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postal tyvek glued down with silicone over 6 1/4 inch holes is whats under the two coffee filters that are rubberbanded down over the lids as dust covers
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NEWT TESTED, NEWT APPROVED.
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taGyo
Strainiac/AMU



Registered: 10/16/14
Posts: 18,802
Loc: Journal Land
Last seen: 5 years, 3 months
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Re: first g2g transfer success? [Re: globejs]
#22505789 - 11/10/15 04:05 PM (8 years, 2 months ago) |
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Well if you have the tyvek I'd take off the coffee filters, they're not doing anything for you at that point 
-------------------- Gyo's Better Grows TNF Q&A AMU Q&A Dominus fortunae meae sum
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FungusOfTheBungus


Registered: 02/05/15
Posts: 121
Last seen: 7 years, 8 months
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Re: first g2g transfer success? [Re: globejs]
#22505849 - 11/10/15 04:18 PM (8 years, 2 months ago) |
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There's not really any need to warm the jars to 84 F. I let mine colonize in room temperatures. Warmer temps just encourage contamination growth.
You might want to shake your jars. That will spread the colonized kernels/grains to the other parts of the jar and increase colonization speed.
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spacechildo
proletarians rise up


Registered: 01/24/13
Posts: 19,243
Loc: Babylon
Last seen: 6 years, 4 months
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globejs
lollygagger



Registered: 08/18/07
Posts: 414
Last seen: 3 months, 21 days
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Re: first g2g transfer success? [Re: spacechildo]
#22506104 - 11/10/15 05:04 PM (8 years, 2 months ago) |
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it doesnt hurt to have more layers between contamination and the grains, it gives me peace of mind so its worth it. and my jars were growing very slowly, and they have since started perking up since they have warmed up. if the contamination is in there its in there right? being hotter isn't going to make them go south. the jars sat for almost a month before use with no contamination so i know there's nothing in them, besides whatever i put in them during the transfer.
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NEWT TESTED, NEWT APPROVED.
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spacechildo
proletarians rise up


Registered: 01/24/13
Posts: 19,243
Loc: Babylon
Last seen: 6 years, 4 months
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Re: first g2g transfer success? [Re: globejs]
#22506119 - 11/10/15 05:09 PM (8 years, 2 months ago) |
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higher temps leads to condensation and higher temps + moisture is a breeding ground for bacteria. so yeah its a problem. 80F is absolute max, 70-75 room temp is perfect.
grains have endospores, tiny things you cant see with the bare eye. they dont die in the pc run they are just knocked back.
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globejs
lollygagger



Registered: 08/18/07
Posts: 414
Last seen: 3 months, 21 days
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Re: first g2g transfer success? [Re: globejs]
#22506122 - 11/10/15 05:10 PM (8 years, 2 months ago) |
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i also know to shake the jars, i did with two right after the transfer, and one i didnt to test a theory i had about spreading contamination as this was my first g2g transfer and i wanted to test some stuff out.
I was waiting to see if any contamination showed up and let them colonize a bit before shaking them up again
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NEWT TESTED, NEWT APPROVED.
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mushpunx
Fungus Punk



Registered: 04/20/14
Posts: 13,394
Last seen: 11 days, 11 hours
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Re: first g2g transfer success? [Re: globejs]
#22506196 - 11/10/15 05:26 PM (8 years, 2 months ago) |
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"One jar was not shaken after a one inch layer of rye was added to the top in hopes that if contam was introduced, the mycelium would overtake it by growing over it downward rather than shaken into the jar anywhere where the mycelium would not be near it to overtake it. Is that bad logic?"
Yes, bad logic haha.
84 degrees is too hot man. You can away with a little bacteria in your spawn and have a successful grow but if you encourage it with high temps the bacteria can take right over. If your room temp is at least 65 lose the incubator, it will do more harm than good. 70s is perfect for spawn.
Case in point, I had a round of spawn inoculated with clean culture on agar. Left town for 4 days during hot part of summer, air conditioner quit. All spawn was over taken by bacteria.
Take the coffee filters off. If you have a proper GE filter there is no reason for it, it might even restrict GE a little (maybe).
To answer your question, it is rare that you see mold in spawn jars. Usually molds like trich will hide in grain spawn through inoculation , G2G and colonization of substrate. The green molds usually show up during fruiting, as you are in essence fruiting the mold as well. Bacteria might be visible in grain spawn through sight or odor.
This is why it is imperative to inoculate your master jars with clean culture on agar. If you arent using agar you might as well inoculate all jars with spores solution rather than risk G2G.
So here are my thoughts.
1. If room temp is over 65 F then lose the incubator.
2. Lose the coffee filters if you have proper GE lids.
3. inoculate master jars with clean culture on agar, as spore syringes are intended for the more forgiving BRF PF type jars, rarely ever clean enough to be put directly to grain. If you cant be sure the master jar was inoculated with clean culture, you will spread whatever to recieving jars.
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 Amateur Mycologists United AMU Q&A
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mushpunx
Fungus Punk



Registered: 04/20/14
Posts: 13,394
Last seen: 11 days, 11 hours
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Re: first g2g transfer success? [Re: mushpunx]
#22506228 - 11/10/15 05:34 PM (8 years, 2 months ago) |
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You are gunna make yourself miserable spraying all the walls and air down in a small plastic tent like that. How big is the room the flow hood in? You only work in the sterile flow of the hood, you can probably lose the plastic.
Set up the hood on a table high off the ground, try to put a sheet of glass in front of the hood as its easy to wipe clean.
Spray the face of the filter with lysol and let sit for a minute.Wipe the table in front of the hood down. Run hood for an hour or two before you start work to scrub the air.
Shower, don clean clothes from the wash, clean tyvek sleeves, face mask and gloves. Wipe gloves with ISO between work. Anything that goes in front of hood should be wiped thoroughly with ISO and especially the rim of jars.Tools flame sterilized.
Never move a hand or anything between the sterile flow and open work.
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 Amateur Mycologists United AMU Q&A
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