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Offlinefreemynd
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question before inoculation
    #7093907 - 06/26/07 09:48 AM (13 years, 10 months ago)

If jars were sterilized with tape over holes AND foil with a rubberband over that. Is it best to remove foil and tape before inoculating? Or best to just poke right through the foil and tape? Also, if one can remove the foil and tape, how long must one wait?


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InvisibleGoodbyeOrb
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Re: question before inoculation [Re: freemynd]
    #7093933 - 06/26/07 10:08 AM (13 years, 10 months ago)

eh, remove them before innoc. If you have dry verm layer, don't even bother putting them back afterwards.


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Offlinefreemynd
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Re: question before inoculation [Re: GoodbyeOrb]
    #7093966 - 06/26/07 10:26 AM (13 years, 10 months ago)

Quote:

GoodbyeOrb said:
eh, remove them before innoc. If you have dry verm layer, don't even bother putting them back afterwards.




Ok, is it recommended to do this inside the glovebox? Or is it better to do it outside? Do gloves need to be worn, etc?


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Re: question before inoculation [Re: freemynd]
    #7093985 - 06/26/07 10:37 AM (13 years, 10 months ago)

If you are using PF jars with a verm filter, it doesn't really matter.  HOWEVER, if you have a glove box, why not use it, and ALWAYS wear gloves. :thumbup:


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Re: question before inoculation [Re: GoodbyeOrb]
    #7094013 - 06/26/07 10:49 AM (13 years, 10 months ago)

Are we talking grain jars or cakes,and as far as the glove box I say do everything in it aside from flaming needles


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Offlinefreemynd
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Re: question before inoculation [Re: Blutjager]
    #7094025 - 06/26/07 10:54 AM (13 years, 10 months ago)

Quote:

Blutjager said:
Are we talking grain jars or cakes,and as far as the glove box I say do everything in it aside from flaming needles




PF BRF cakes ...with dry Verm layer on top. I'll tell him to do it inside the glovebox WITH gloves. Another thing he noticed. When he uses a regular lighter to sterilize it seems to take FOREVER to get the tip of the needle red, and when it does, it's barely red. Is this good enough for sterilization?


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Re: question before inoculation [Re: freemynd]
    #7094080 - 06/26/07 11:23 AM (13 years, 10 months ago)

Yea its good enough,half the time I don't even flame needles anymore and just keep them with an alcohol pad wrapped around them,they are cheap as hell at the pharmacy and individually wrapped for people who need to give them self injections of medicine.Pic up a box and you will be good as gold for a while:thumbup:


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Re: question before inoculation [Re: freemynd]
    #7094106 - 06/26/07 11:33 AM (13 years, 10 months ago)

I've gotten this one before.  What you need to do is move the needle closer to the tip of the flame.  Also, as a side note, for a cheap and easy alcohol flame, take a small votive candle, or any other small dish, fill with rubbing alcohol, and light on fire.  Works wonders.  You could even use a shot glass.  Wise man say however, do not use alcohol lamp in glovebox full of lysol fumes.  Old Chinese proverb. :thumbup:


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Offlinefreemynd
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Re: question before inoculation [Re: Blutjager]
    #7094116 - 06/26/07 11:37 AM (13 years, 10 months ago)

ok, good tip. I think he usually wipes the needles down with a paper towl soaked in rubbing alcohol, then flame sterilizes. here is a picture of his glovebox. it looks dirty because he washed it with bleach spray and it left streaks. also, the holes were hard to cut, that's why they look so jagged...lol. what do you think?



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Re: question before inoculation [Re: freemynd]
    #7094751 - 06/26/07 03:04 PM (13 years, 10 months ago)

Personally, I keep the foil and everything on until it's in the glovebox (yes use one!) Then I remove the foil and innoc thru the tape. The hole made by the tape is enough for gas exchange.

Blut you can get sterile needles at the pharmacy? Hell I never thought to check, just figured you had to have a scrip or something.


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Offlinefreemynd
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Re: question before inoculation [Re: mushbaby]
    #7095163 - 06/26/07 04:46 PM (13 years, 10 months ago)

Quote:

mushbaby said:
Personally, I keep the foil and everything on until it's in the glovebox (yes use one!) Then I remove the foil and innoc thru the tape. The hole made by the tape is enough for gas exchange.




That's a good idea. I'll tell my friend not to even bother trying to take that sticky tape off. I think the mistake he made last time was he removed the foil and tape before he threw it in the glovebox. And he probably ended up touching the holes. He also noticed it was extremely difficult to move around in his glovebox. Maybe it's too small.


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Offlinefiggusfiddus
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Re: question before inoculation [Re: freemynd]
    #7095287 - 06/26/07 05:16 PM (13 years, 10 months ago)

I use some pore tape over the holes. It's easy enough to put on, and it seems to keep large particles out so that no huge quantity of contamination is introduced after inoculation. I've never seen in vitro mold, and I plan not to, this way.

But eh, make sure you use clean, preferably gloved hands when you put the tape on. To get a little piece of tape off, you pretty much have to touch your skin to the sticky part. Needless to say, sticky stuff tends to GRAB stuff from your fingertips... including bacteria, mold spores, et cetera. And then by putting the tape over the holes you've basically held these contaminants face-to-face with your verm layer.


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Re: question before inoculation [Re: figgusfiddus]
    #7095310 - 06/26/07 05:21 PM (13 years, 10 months ago)

Quote:

figgusfiddus said:


But eh, make sure you use clean, preferably gloved hands when you put the tape on. To get a little piece of tape off, you pretty much have to touch your skin to the sticky part. Needless to say, sticky stuff tends to GRAB stuff from your fingertips... including bacteria, mold spores, et cetera. And then by putting the tape over the holes you've basically held these contaminants face-to-face with your verm layer.




Ah crap, I don't think he washed his hands very good while preparing the verm/BRF and applying the tape. I guess he figured it'd sterilize during the PC. Should he dump out his jars and start over?


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Re: question before inoculation [Re: freemynd]
    #7095372 - 06/26/07 05:40 PM (13 years, 10 months ago)

Jeez, no. It's fine.

Don't dump them until they're actually contaminated. Hand-washing is one of those precautionary things, that's all. It's not a make-or-break situation whatsoever.

Wait, was the tape applied prior to PCing? If so, you don't have to worry. I apply mine afterward, because poking an inoculation hole in pore tape tends to break through it. Also, my pore tape melts before 255F. Really, the tape isn't even 100% necessary--that's what the prophylactic verm layer is for.

Hand-washing is totally unnecessary when mixing your substrate. The PC *will* take care of that. I do mine in the kitchen, next to the trash, with unwashed hands and a wooden spoon, which are all terrible for sterile conditions. It's totally fine, that is indeed why we sterilize it.


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Offlinefreemynd
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Re: question before inoculation [Re: figgusfiddus]
    #7095652 - 06/26/07 06:38 PM (13 years, 10 months ago)

Oh phew....yeah, the tape was applied before PCing. So it looks like everything is good. He'll remove the foil (inside the glovebox) and insert needle through tape...and leave alone. Sounds like a great plan!!


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