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mthawkins
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Questions regarding inoculation and still air box (WBS/Franks Tek)
#18719585 - 08/17/13 04:35 PM (10 years, 5 months ago) |
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I've made my still air box by turning holes into it.. Now it's time to inoculate. My goods are Clorox bleach spray, 70% iso alch, and a lighter. I assume the process will go as such: I will spray the bleach into the still air box, then give it a good run down. Wait 10-15 minutes then place the jars into the box after removing foil. (Should I also rub down the jars?)
I will then first burn the needle till red (inside or outside box?) , then run down with alch with cotton ball. Inoculate 1cc into the RTV port. Should I press the needle into the seeds or just drop above it? I assume I clean the gloves with the rubbing alcohol as well. I just don't want to catch on fire. Do I burn needle inside chamber? Repeat repeat
-------------------- “Never laugh at live dragons.” - J.R.R Tolkien
Edited by mthawkins (08/17/13 04:37 PM)
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blojo02184
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Re: Questions regarding inoculation and still air box (WBS/Franks Tek) [Re: mthawkins]
#18719614 - 08/17/13 04:41 PM (10 years, 5 months ago) |
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1. wiping jars optional, just not thetop. 2. flame red hot outside of box. Any alcohol vapors could explode. 3. running down tye glass is the best option.
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mthawkins
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Re: Questions regarding inoculation and still air box (WBS/Franks Tek) [Re: mthawkins]
#18719616 - 08/17/13 04:43 PM (10 years, 5 months ago) |
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I also have disinfectant wipes from Lysol and Clorox
.. Ok, so ill drop it above the seeds to let run down.
-------------------- “Never laugh at live dragons.” - J.R.R Tolkien
Edited by mthawkins (08/17/13 04:43 PM)
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mthawkins
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Re: Questions regarding inoculation and still air box (WBS/Franks Tek) [Re: mthawkins]
#18719650 - 08/17/13 04:53 PM (10 years, 5 months ago) |
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I wonder if rubbing some Lysol on my arms would be just as good as sleeves ( while sti wearing gloves)
-------------------- “Never laugh at live dragons.” - J.R.R Tolkien
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Pestile

Registered: 05/02/13
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Re: Questions regarding inoculation and still air box (WBS/Franks Tek) [Re: mthawkins]
#18719665 - 08/17/13 04:56 PM (10 years, 5 months ago) |
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Quote:
mthawkins said: I wonder if rubbing some Lysol on my arms would be just as good as sleeves ( while sti wearing gloves)
Nah, there'll still be dry skin flakes and other stuff.
--------------------
   The Corbett Report Open Source Intelligence News
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BloodKil
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Re: Questions regarding inoculation and still air box (WBS/Franks Tek) [Re: mthawkins]
#18719677 - 08/17/13 04:58 PM (10 years, 5 months ago) |
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Don't wipe down the needle with alcohol after flaming... just flame, inject needle, wait a couple seconds, shoot spore solution.
You honestly don't even need to wait, as the first bit of solution through the needle will cool it, but when I was working with needles I liked to wait a few seconds after inserting the needle to give a better chance at more viable spores entering.
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mthawkins
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Re: Questions regarding inoculation and still air box (WBS/Franks Tek) [Re: BloodKil]
#18719694 - 08/17/13 05:03 PM (10 years, 5 months ago) |
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How about putting tape around my arm? Or Lysol a bag and wrap it ?
I don't put the needle into the seeds, correct? Just let it drop along the side.. Or middle? I'm about to do it once I get a good response
I've already sprayed then wiped the container down with Lysol... Rubbed the jars down with Lysol, sprayed in the air, removed foil and placed jars into container
-------------------- “Never laugh at live dragons.” - J.R.R Tolkien
Edited by mthawkins (08/17/13 05:05 PM)
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K1ngSp4de
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Re: Questions regarding inoculation and still air box (WBS/Franks Tek) [Re: BloodKil]
#18719698 - 08/17/13 05:05 PM (10 years, 5 months ago) |
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Always clean your jars with alcohol before placing in the sab, then wipe the top off again right before inoculation.
Inoculate toward the edge of the jar, don't touch the grain with the needle. This way you will be able to tell when it germinates, and watch progress.
-------------------- PC Repair and Troubleshooting Forum If a law is unjust, a man is not only right to disobey it, he is obligated to do so. - Thomas Jefferson Si peccasse negamus fallimur et nulla est in nobis veritas.
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mthawkins
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Re: Questions regarding inoculation and still air box (WBS/Franks Tek) [Re: K1ngSp4de]
#18719725 - 08/17/13 05:12 PM (10 years, 5 months ago) |
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How about gloves? Clean with alch too? Bag or tape Around my arms?
I'm cleaning bottle with alch rub and around the cotton /rtv as we speak. I've lysoled the container, no alch tho
-------------------- “Never laugh at live dragons.” - J.R.R Tolkien
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K1ngSp4de
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Re: Questions regarding inoculation and still air box (WBS/Franks Tek) [Re: mthawkins]
#18719750 - 08/17/13 05:19 PM (10 years, 5 months ago) |
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Yhea you have to be as clean a possible, rub alcohol all over your gloved hands and your arms. Don't worry about wrapping your arms in anything, just rub them in alcohol every like 4-5 jars. I keep a saturated paper towel on the bottom of the sab for wiping the jar lids and my gloves. If you're doing it right your arms shouldn't really matter as they should never be above or touch anything.
-------------------- PC Repair and Troubleshooting Forum If a law is unjust, a man is not only right to disobey it, he is obligated to do so. - Thomas Jefferson Si peccasse negamus fallimur et nulla est in nobis veritas.
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mthawkins
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Re: Questions regarding inoculation and still air box (WBS/Franks Tek) [Re: K1ngSp4de]
#18719866 - 08/17/13 05:51 PM (10 years, 5 months ago) |
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What I ended up doing was cleaning the tub as much as possible, wiped down the jars and lids. Put gloves and bag around my arm wrapped with duck tape. Cleaned it with alch. I inoculated to let drip down side of jar. 1cc each. I used the whole syringe. After ever inoculation, I would light a fire on the needle then wipe with alch, along with the rtv injection port. Rinse and repeat. What do I do with them now? Should I just place in my kitchen cabinet ?
-------------------- “Never laugh at live dragons.” - J.R.R Tolkien
Edited by mthawkins (08/17/13 05:58 PM)
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K1ngSp4de
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Re: Questions regarding inoculation and still air box (WBS/Franks Tek) [Re: mthawkins]
#18719897 - 08/17/13 06:02 PM (10 years, 5 months ago) |
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You should NOT have wiped the needle with alcohol, in fact someone else already told you. Its counter productive, the needle is as sterile as its gonna get after flaming.
-------------------- PC Repair and Troubleshooting Forum If a law is unjust, a man is not only right to disobey it, he is obligated to do so. - Thomas Jefferson Si peccasse negamus fallimur et nulla est in nobis veritas.
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mthawkins
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Re: Questions regarding inoculation and still air box (WBS/Franks Tek) [Re: K1ngSp4de]
#18719923 - 08/17/13 06:09 PM (10 years, 5 months ago) |
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Will that cause contamination? I've read everywhere to wipe with alcohol.. Guess ill see :/
-------------------- “Never laugh at live dragons.” - J.R.R Tolkien
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K1ngSp4de
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Re: Questions regarding inoculation and still air box (WBS/Franks Tek) [Re: mthawkins]
#18719933 - 08/17/13 06:11 PM (10 years, 5 months ago) |
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Yes it can. Then everywhere you've been is wrong ask any TC.
-------------------- PC Repair and Troubleshooting Forum If a law is unjust, a man is not only right to disobey it, he is obligated to do so. - Thomas Jefferson Si peccasse negamus fallimur et nulla est in nobis veritas.
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Pestile

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Re: Questions regarding inoculation and still air box (WBS/Franks Tek) [Re: mthawkins]
#18721942 - 08/18/13 07:48 AM (10 years, 5 months ago) |
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Quote:
mthawkins said: Will that cause contamination? I've read everywhere to wipe with alcohol.. Guess ill see :/
Quote:
Notahacker420 said: Wiping alcohol on a flame sterilized needle is like wiping shit on your face after a shower.
Quote:
RR
Wiping with alcohol between jars is a good way to introduce mold spores into each jar, always flame sterilize between jars.
How are people who have been here this long still giving shit advice?
http://www.shroomery.org/forums/showflat.php/Number/17737262#17737262
--------------------
   The Corbett Report Open Source Intelligence News
Edited by Pestile (08/18/13 07:50 AM)
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spacechildo
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Re: Questions regarding inoculation and still air box (WBS/Franks Tek) [Re: Pestile]
#18722002 - 08/18/13 08:41 AM (10 years, 5 months ago) |
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Most threads I've read where someone tells you to wipe your sterilized needle with only sanitizing alcohol, someone has chimed in and said NOT to do it. Kinnda like this thread. People still do it.
Tip to you op; check the dates on the threads you read. Lots of info has changed over the years, I usually search for threads from the last 2 years.
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Nakor420
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Re: Questions regarding inoculation and still air box (WBS/Franks Tek) [Re: spacechildo]
#18722026 - 08/18/13 08:56 AM (10 years, 5 months ago) |
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Well, I have a large paper towel soaked with 91% iso when I flame sterilize, I use the pad to cool the needle...here is why...one, I have a flow hood, so I know the pad is sterile...two, cooling the needle by pushing solution through it kills some of your spores...by doing this I get roughly two more inoculations from each syringe..though I have to agree that it's not a good idea if you don't have laminar flow.
Edited by Nakor420 (08/18/13 08:57 AM)
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tbagtag
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Re: Questions regarding inoculation and still air box (WBS/Franks Tek) [Re: Nakor420]
#18722050 - 08/18/13 09:08 AM (10 years, 5 months ago) |
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Quote:
Nakor420 said: Well, I have a large paper towel soaked with 91% iso when I flame sterilize, I use the pad to cool the needle...here is why...one, I have a flow hood, so I know the pad is sterile...two, cooling the needle by pushing solution through it kills some of your spores...by doing this I get roughly two more inoculations from each syringe..though I have to agree that it's not a good idea if you don't have laminar flow.
It's like putting a condom on after you came in the girl, pointless. With how hot that needle is nothing is going to grow on it. Until you cool it down rapidly on organic matter which I'm assuming is sitting in a bulk pack, or kitchen, or bathroom. Remember you can grow mold on a paper towel, unless your autoclaving your paper towels all you've been doing is getting lucky.
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Leon76


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Re: Questions regarding inoculation and still air box (WBS/Franks Tek) [Re: tbagtag]
#18722061 - 08/18/13 09:15 AM (10 years, 5 months ago) |
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Mold won't grow on alcohol. As long as the needle only came in contact with the alcohol soaked part of the paper towel it would be fine. But, no point after heated.
-------------------- “I have absolutely no pleasure in the stimulants in which I sometimes so madly indulge. It has not been in the pursuit of pleasure that I have periled life and reputation and reason. It has been the desperate attempt to escape from torturing memories, from a sense of insupportable loneliness and a dread of some strange impending doom.” ― Edgar Allan Poe
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Nakor420
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Re: Questions regarding inoculation and still air box (WBS/Franks Tek) [Re: tbagtag]
#18722142 - 08/18/13 09:46 AM (10 years, 5 months ago) |
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Quote:
tbagtag said:
Quote:
Nakor420 said: Well, I have a large paper towel soaked with 91% iso when I flame sterilize, I use the pad to cool the needle...here is why...one, I have a flow hood, so I know the pad is sterile...two, cooling the needle by pushing solution through it kills some of your spores...by doing this I get roughly two more inoculations from each syringe..though I have to agree that it's not a good idea if you don't have laminar flow.
It's like putting a condom on after you came in the girl, pointless. With how hot that needle is nothing is going to grow on it. Until you cool it down rapidly on organic matter which I'm assuming is sitting in a bulk pack, or kitchen, or bathroom. Remember you can grow mold on a paper towel, unless your autoclaving your paper towels all you've been doing is getting lucky.
I think you missed the entire point of what I said,lol... you aren't telling me anything I don't know. lol
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Nakor420
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Re: Questions regarding inoculation and still air box (WBS/Franks Tek) [Re: Leon76]
#18722148 - 08/18/13 09:47 AM (10 years, 5 months ago) |
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Quote:
Leon76 said: Mold won't grow on alcohol. As long as the needle only came in contact with the alcohol soaked part of the paper towel it would be fine. But, no point after heated.
Except the point I just made about getting more inoculations from a syringe by cooling the needle before you push solution through it.
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coffeehead
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Re: Questions regarding inoculation and still air box (WBS/Franks Tek) [Re: Leon76]
#18722165 - 08/18/13 09:53 AM (10 years, 5 months ago) |
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I work with a glove box inside a still air room that I clean the best I can. I prepare the box by wiping it down with Clorox wipes and spraying some Lysol and Clorox water inside. As long as you don't overdo it with the Lysol, the Clorox spray will make it safe to use the lighter inside of the box. I also use Tyvek sleeves that I've sprayed with Lysol. I put the jars straight into the box from the PC. And I stopped using foil because water inevitably condensed on it and got onto my filter disks. I also spray down MY PC with Lysol and Clorox water before putting it in my clean room since it has sat on the stove overnight in my messy kitchen. I use a face mask and spray my gloves with Lysol and then Clorox water. I use Lysol wipes liberally on anything else that looks like it may pose a contamination risk.
Zero contaminations experienced using this method.
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FrankHorrigan
The Inquisition



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Re: Questions regarding inoculation and still air box (WBS/Franks Tek) [Re: coffeehead]
#18722400 - 08/18/13 10:59 AM (10 years, 5 months ago) |
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This thread is out of hand 
Don't wipe your flame sterilized tools with alcohol.
Either cool them by making cuts in agar (scalpel or loop) or squirting a few drops from the syringe.
Yes, you can get away with it. But fire kills things far better than alcohol so there really is no reason to risk it 
Quote:
blojo02184 said: 1. wiping jars optional, just not thetop. 2. flame red hot outside of box. Any alcohol vapors could explode. 3. running down tye glass is the best option.

Don't worry so much, the main issues you will have with the SAB is drafts in the room and your movements. It takes some practice.
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Nakor420
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Re: Questions regarding inoculation and still air box (WBS/Franks Tek) [Re: FrankHorrigan]
#18722432 - 08/18/13 11:10 AM (10 years, 5 months ago) |
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Quote:
FrankHorrigan said: This thread is out of hand 
Don't wipe your flame sterilized tools with alcohol.
Either cool them by making cuts in agar (scalpel or loop) or squirting a few drops from the syringe.
Yes, you can get away with it. But fire kills things far better than alcohol so there really is no reason to risk it 
Quote:
blojo02184 said: 1. wiping jars optional, just not thetop. 2. flame red hot outside of box. Any alcohol vapors could explode. 3. running down tye glass is the best option.

Don't worry so much, the main issues you will have with the SAB is drafts in the room and your movements. It takes some practice.
Frank, I flame sterlize, but you don't think cooling before squirting is worth it for the extra spores that survive the needle? I mean with my flow hood, I don't see the downside at all, just the two grain jars I started adding to my MS's.
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FrankHorrigan
The Inquisition



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Re: Questions regarding inoculation and still air box (WBS/Franks Tek) [Re: FrankHorrigan]
#18722441 - 08/18/13 11:12 AM (10 years, 5 months ago) |
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The downside is:
Quote:
FrankHorrigan said: fire kills things far better than alcohol so there really is no reason to risk it 
Flowhood or not, nothing is 100% effective. I don't chance it over a couple drops of spores hissing to the floor. You've got a LOT of them, and there are plenty more available 
I like as much success as possible so...
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Nakor420
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Re: Questions regarding inoculation and still air box (WBS/Franks Tek) [Re: FrankHorrigan]
#18722450 - 08/18/13 11:15 AM (10 years, 5 months ago) |
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It's never failed for me and since I started doing it I have literally gotten two more jars per syringe..So this will just have to be one of those things we agree to disagree on. ...Though I would normally bow to your wisdom.
Edited by Nakor420 (08/18/13 11:16 AM)
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FrankHorrigan
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Re: Questions regarding inoculation and still air box (WBS/Franks Tek) [Re: Nakor420]
#18722461 - 08/18/13 11:19 AM (10 years, 5 months ago) |
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I'm saying it's an added risk, not that you have to stop what you're doing 
That is more of a fact than anything. Mold spores are not all killed by alcohol.
I have hundreds of prints, a dozen syringes...spores are cheap and easy to acquire but my time and materials are valuable.
This is of course just my opinion.
If you'd like to get more acquainted with the feeling of "spore overload," send me a
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mthawkins
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Re: Questions regarding inoculation and still air box (WBS/Franks Tek) [Re: FrankHorrigan]
#18722517 - 08/18/13 11:39 AM (10 years, 5 months ago) |
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Gah, now you guys have me thinking I need to purchase another syringe and go at it again lol..
-------------------- “Never laugh at live dragons.” - J.R.R Tolkien
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tbagtag
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Re: Questions regarding inoculation and still air box (WBS/Franks Tek) [Re: FrankHorrigan]
#18722531 - 08/18/13 11:44 AM (10 years, 5 months ago) |
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Quote:
FrankHorrigan said: Mold spores are not all killed by alcohol.
This is 100% fact, I worked where I had to be certified in blood borne and airborne pathogens. Mold's is one of the things discussed as I had to learn to get open areas near operating room clean. Usually its a combination of chemicals like cavacide, matacide, bleach, alcohol and all done in a specific order in a timed sequence to remove all living spore and bacteria.
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Nakor420
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Re: Questions regarding inoculation and still air box (WBS/Franks Tek) [Re: tbagtag]
#18722576 - 08/18/13 12:01 PM (10 years, 5 months ago) |
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Quote:
tbagtag said:
Quote:
FrankHorrigan said: Mold spores are not all killed by alcohol.
This is 100% fact, I worked where I had to be certified in blood borne and airborne pathogens. Mold's is one of the things discussed as I had to learn to get open areas near operating room clean. Usually its a combination of chemicals like cavacide, matacide, bleach, alcohol and all done in a specific order in a timed sequence to remove all living spore and bacteria.
The way I do it, is I soak a paper towel folded in 1/4ths with laminar flow, lots of iso 91%, then after the flame sterilization, I just use a fold in the p towel to cool the needle from the lower outside of the needle, and "sizzle" my way up to ALMOST the tip, so the minute amount of possible mold spores doesn't effect the flow of solution.
 Plus anything that MIGHT be on on the base/middle of the needle will likely be rubbed off by the injection port.
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tbagtag
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Re: Questions regarding inoculation and still air box (WBS/Franks Tek) [Re: Nakor420]
#18722589 - 08/18/13 12:05 PM (10 years, 5 months ago) |
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They are dead when you flame cook them fuckers.
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Nakor420
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Re: Questions regarding inoculation and still air box (WBS/Franks Tek) [Re: tbagtag]
#18722613 - 08/18/13 12:12 PM (10 years, 5 months ago) |
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yeah the ones already in the needle, but if you cool with solution, you are also killing enough to knock up a jar on top of that.
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tbagtag
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Re: Questions regarding inoculation and still air box (WBS/Franks Tek) [Re: Nakor420]
#18722644 - 08/18/13 12:19 PM (10 years, 5 months ago) |
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Take it easy, treat that SHIP like a woman you're about to make sweet love to. After you get it heated up, count to 10 before you stick it in. Wait another 5 seconds and let out a drop or 2, then blow your load. You keep it clean classy and simple.
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Nakor420
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Re: Questions regarding inoculation and still air box (WBS/Franks Tek) [Re: tbagtag]
#18722660 - 08/18/13 12:23 PM (10 years, 5 months ago) |
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yeah, but even after waiting, then putting to my cool pad, I still hear sizzling. This debate is silly,lol. I'm out.
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mthawkins
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Re: Questions regarding inoculation and still air box (WBS/Franks Tek) [Re: Nakor420]
#18722737 - 08/18/13 12:43 PM (10 years, 5 months ago) |
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When should I start seeing contamination vs colonization?
-------------------- “Never laugh at live dragons.” - J.R.R Tolkien
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tbagtag
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Re: Questions regarding inoculation and still air box (WBS/Franks Tek) [Re: mthawkins]
#18722799 - 08/18/13 01:03 PM (10 years, 5 months ago) |
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Depends on the contamination, usually if you see growth within the first 48 hours from w MS syringe it's not a good sign.
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FrankHorrigan
The Inquisition



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Re: Questions regarding inoculation and still air box (WBS/Franks Tek) [Re: Nakor420]
#18724830 - 08/18/13 09:18 PM (10 years, 5 months ago) |
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Quote:
Nakor420 said: This debate is silly
What debate?
Flame kills things better than alcohol. You said you will continue trusting the alcohol over the flame. No one here is trying to stop you, we're just letting you know that it is counterproductive/pointless 
Quote:
FrankHorrigan said: If you'd like to get more acquainted with the feeling of "spore overload," send me a 
You miss this nakor?
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Psilicon
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Re: Questions regarding inoculation and still air box (WBS/Franks Tek) [Re: FrankHorrigan]
#18725440 - 08/18/13 11:41 PM (10 years, 5 months ago) |
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You never responded to my PM on that subject, Frank. Maybe Nakor already did. 
Nakor, if saving spores is an issue for you, with a PC you can either do agar work or go straight to grain and then G2G for thousands of quarts with just a single drop of spore solution.
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FrankHorrigan
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Re: Questions regarding inoculation and still air box (WBS/Franks Tek) [Re: Psilicon]
#18725501 - 08/18/13 11:58 PM (10 years, 5 months ago) |
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Quote:
van der griegen said: You never responded to my PM on that subject, Frank.
Well send me another.
I'm notorious for reading pm's while drunk, forgetting about them, and then deleting them with the rest every couple weeks
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Nakor420
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Re: Questions regarding inoculation and still air box (WBS/Franks Tek) [Re: FrankHorrigan]
#18726117 - 08/19/13 06:15 AM (10 years, 5 months ago) |
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I don't "trust it over flame to sterilize" I'm still sterilizing with my alcohol lamp, lol..... I use my alcohol pad to cool the needle...that's IT.
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