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OfflineCubeSteakPoo
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Registered: 04/27/16
Posts: 95
Last seen: 6 years, 9 months
Sterile Technique Question
    #23979800 - 01/03/17 07:46 PM (7 years, 27 days ago)

Hey guys,

Sorry if this is in the incorrect forum section, but i was having a hard time navigating and thought this would be sufficient enough of a place to ask this.

After some serious contamination  issues form what i beleive to be faulty syringes, Im trying to cover every step for the next time i inoculate.

A lot of the teks say that when you do your sterile technique, that you should use Latex gloves.

This may seem like a dumb question, but if i have "Non-Sterile Vinyl Gloves" I should have no trouble disenfecting them with isopropyl alcohol, and be good to go? or is there something specifically neccessary with latex gloves?


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OfflineTira
I'm a teapot


Registered: 11/20/10
Posts: 1,202
Loc: Turkey
Last seen: 6 years, 3 months
Re: Sterile Technique Question [Re: CubeSteakPoo]
    #23980444 - 01/04/17 01:35 AM (7 years, 26 days ago)

Those will do fine, keep in mind that alcohol should be 70% and not 90%


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OfflineRikazi
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Registered: 10/13/16
Posts: 60
Last seen: 3 months, 6 days
Re: Sterile Technique Question [Re: Tira]
    #23980491 - 01/04/17 02:22 AM (7 years, 26 days ago)

why not 90%?


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OfflineTira
I'm a teapot


Registered: 11/20/10
Posts: 1,202
Loc: Turkey
Last seen: 6 years, 3 months
Re: Sterile Technique Question [Re: Rikazi]
    #23980648 - 01/04/17 06:26 AM (7 years, 26 days ago)

Quote:

An ethanol percentage of 50-80% destroys the cell wall/membrane of bacteria by denaturing their proteins and dissolving their lipids (effective against most bacteria, fungi and some viruses; ineffective against bacterial spores). Therefore, the ethanol has to pass the bacterial membrane/wall to get into the bacteria - if you use 100% ethanol instead, the bacteria get 'sealed' and they will survive...




Source


--------------------
Useful Links for Beginners

The Basics              AMU Teks

Frank''s Teks          Agar

Noob Forum Reccomended Teks

Agar for guaranteed spawn,
Proper pasteurization for guaranteed substrate.


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OfflineRolledUhhp
Amateur Cultivator

Registered: 08/10/16
Posts: 246
Last seen: 3 years, 6 months
Re: Sterile Technique Question [Re: Tira]
    #23982349 - 01/04/17 07:33 PM (7 years, 26 days ago)

Quote:

Tira said:
Quote:

An ethanol percentage of 50-80% destroys the cell wall/membrane of bacteria by denaturing their proteins and dissolving their lipids (effective against most bacteria, fungi and some viruses; ineffective against bacterial spores). Therefore, the ethanol has to pass the bacterial membrane/wall to get into the bacteria - if you use 100% ethanol instead, the bacteria get 'sealed' and they will survive...




Source





Nice, thank you!


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OfflineCrispykoot
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Registered: 10/16/16
Posts: 5,921
Loc: Flag
Last seen: 18 hours, 7 minutes
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Re: Sterile Technique Question [Re: RolledUhhp]
    #23990978 - 01/07/17 09:15 PM (7 years, 23 days ago)

This has the specs for Isopropyl, the above refers to ethanol: Chemical Disinfectants
Alcohol
Overview.  In the healthcare setting, "alcohol" refers to two water-soluble chemical compounds—ethyl alcohol and isopropyl alcohol—that have generally underrated germicidal characteristics 482.  FDA has not cleared any liquid chemical sterilant or high-level disinfectant with alcohol as the main active ingredient. These alcohols are rapidly bactericidal rather than bacteriostatic against vegetative forms of bacteria; they also are tuberculocidal, fungicidal, and virucidal but do not destroy bacterial spores. Their cidal activity drops sharply when diluted below 50% concentration, and the optimum bactericidal concentration is 60%–90% solutions in water (volume/volume) 483, 484.

Source: https://www.cdc.gov/hicpac/disinfection_sterilization/6_0disinfection.html


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OfflinePsilocyBen17
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Registered: 10/20/13
Posts: 3,751
Last seen: 1 year, 1 month
Re: Sterile Technique Question [Re: Crispykoot]
    #23991123 - 01/07/17 10:05 PM (7 years, 23 days ago)

IME gloves have never made much of a difference. Before innoculating I wash my hands and forearms with hot water and lots of soap. Then I brush my teeth and gargle with mouthwash.  Then I put on a facemask so Im not breathing directly on my work space. Next I wipe down my bench and the space infront of my flowhood with lysol. Then 70% ISO (in this case using lysol is probably redundant, but I like to think it saves on alcohol, i hate when running out prevents me from starting a project).

I choose not to wear gloves anymore because I have about a 98% success rate and I never found that wearing gloves made any difference. I did notice that gloves caused me to drop my scalpel and fumble with jar lids, which is bad technique. Besides all this they were hot and uncomfortable.

If you are using a SAB instead of a bench just wipe it down with hot, soapy water, load it up, and mist the air to arrest the contams before you start your work. Its been a few years since I used a SAB but I don't think people wipe the box interior with ISO, only their tools, hands, and work surface. Hopefully someone can correct me if I'm wrong as I plan to use a SAB pretty soon for a class I'm teaching LOL.


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