Home | Community | Message Board

Avalon Magic Plants
This site includes paid links. Please support our sponsors.


Welcome to the Shroomery Message Board! You are experiencing a small sample of what the site has to offer. Please login or register to post messages and view our exclusive members-only content. You'll gain access to additional forums, file attachments, board customizations, encrypted private messages, and much more!

Shop: North Spore North Spore Mushroom Grow Kits & Cultivation Supplies   Left Coast Kratom Buy Kratom Extract   Original Sensible Seeds Bulk Cannabis Seeds   PhytoExtractum Kratom Powder for Sale   Mushroom-Hut Substrate Bags   Myyco.com Isolated Cubensis Liquid Culture For Sale

Jump to first unread post Pages: < Back | 1 | 2  [ show all ]
Some of these posts are very old and might contain outdated information. You may wish to search for newer posts instead.
Offlinekrypto2000
Unknown


Registered: 12/05/06
Posts: 11,579
Last seen: 4 years, 5 months
Re: Sterilizing air of glovebox? [Re: Juiceh]
    #18827275 - 09/11/13 11:01 AM (10 years, 6 months ago)

I've found the best way to sterilize my SAB is to make a bigger SAB and sterilize that inside a yet bigger SAB and then I work in the SAB in the SAB to sterilize the SAB.

Extras: Filter Print Post Top
Offlinemagnesium
Stranger
 User Gallery

Registered: 08/08/13
Posts: 31
Last seen: 9 years, 10 months
Re: Sterilizing air of glovebox? [Re: krypto2000]
    #18840845 - 09/14/13 11:46 AM (10 years, 6 months ago)

I turn the SAB upside down ontop of a towel soaked in bleach, instead of a lid. Feels more comfortable that way.

Extras: Filter Print Post Top
Invisiblefastfred
Old Hand
Male User Gallery

Folding@home Statistics
Registered: 05/17/04
Posts: 6,899
Loc: Dark side of the moon
Re: Sterilizing air of glovebox? [Re: micro]
    #18864901 - 09/19/13 08:11 PM (10 years, 6 months ago)

Quote:

micro said:
Quote:

Notahacker420 said:
Not a lot of people own Bunsen burners, nor is one needed if you have a still air box and good sterile technique.

When you use a bunsen burner, yes everything rises with the heat, but then it falls.




It then goes into the flame because of lower air density, right?

I mean, this has been done for decades but it's a pain unless you are doing something trivial.




I don't see what's so hard about turning on the valve to your $12 bunsen and flicking a lighter.

Dirty air is sucked into the bottom of the bunsen.  Any contams burn up, rise until the air cools then fall down.  Most of the air does not go through the bunsen, but the air is flowing upwards in an area around the bunsen.

In any case, it's standard micro technique.  Nobody in micro uses a SAB and most don't have flowhoods either.


--------------------
It drinks the alcohol and abstains from the weed or else it gets the hose again. -Chemy

The difference between the substances doesn't matter. This is a war on consciousness, on our right to the very essence of what we are. With no control over that, we have no need to speak of freedom or a free society. -fireseed

"If we are going to have a war on marijuana, the least we can do is pull the sick and the dying off the battlefield." -Neal Levine (MPP)

I find the whole "my drug should be legal but yours should be illegal" mindset disgusting and hypocritical. It's what George Bush and company do when they drink a cocktail and debate the best way to imprison marijuana users. -Diploid

Extras: Filter Print Post Top
Offlinekrypto2000
Unknown


Registered: 12/05/06
Posts: 11,579
Last seen: 4 years, 5 months
Re: Sterilizing air of glovebox? [Re: fastfred]
    #18864996 - 09/19/13 08:27 PM (10 years, 6 months ago)

The hole in that diagram looks to be rather low, wouldn't it pull dirty air strait across the petri in that case or is the diagram just off?

Extras: Filter Print Post Top
Offlinecubenpete
Aminita good excuse
Male User Gallery


Registered: 03/13/12
Posts: 837
Loc: Kentucky
Last seen: 2 years, 5 months
Re: Sterilizing air of glovebox? [Re: krypto2000]
    #18865528 - 09/19/13 10:36 PM (10 years, 6 months ago)

I use Oust or Nuetra Air both inside and outside the box and give it 10-15min to settle inside onto 70/90% iso alcohol sprayed/soaked paper towels. Of course I turn off any artificial air flow in my home. Granted I have only run 20+ tubs and 60 petri dishes (kinda new to agar), and have had zero contams in my projects. I usually shower/scrub, mouthwash/brush and wear facemask/gloves.

I use the same procedure when spawning to subs and have had equal success. Fingers crossed!

This does not mean that I do not want the space and maneuverability that a flow hood provides, but doubling the size of my SAB (68qts.. yay!), has made things much less arduous.

Extras: Filter Print Post Top
Invisiblefastfred
Old Hand
Male User Gallery

Folding@home Statistics
Registered: 05/17/04
Posts: 6,899
Loc: Dark side of the moon
Re: Sterilizing air of glovebox? [Re: krypto2000]
    #18867744 - 09/20/13 01:22 PM (10 years, 6 months ago)

Quote:

krypto2000 said:
The hole in that diagram looks to be rather low, wouldn't it pull dirty air strait across the petri in that case or is the diagram just off?




The diagram isn't the best.  Air does get sucked into that hole, but the diagram doesn't really show the larger pattern of air entrainment and convection very well.

The portion sucked into the hole is not nearly as much as the air that is entrained by the rising air from the flame.  The main thing is that the air in the sterile area is fairly clean and moving upwards.  No contams can fall down into your dish in that area.

Explanations often fail, the important fact is that this is the basis of microbiology.  It's been proven to work for hundreds of years and is still how almost all microbiology is done today.  No fancy flow hoods or SABs.  Those can and do work, but they aren't as effective as good sterile technique with a bunsen burner.


-FF

Extras: Filter Print Post Top
OfflinePussyFart
Retired Cultivation Extrodinaire
Male


Registered: 04/08/12
Posts: 22,502
Loc: Orbiting Earth
Last seen: 2 months, 14 days
Trusted Cultivator
Re: Sterilizing air of glovebox? [Re: fastfred]
    #18867950 - 09/20/13 02:13 PM (10 years, 6 months ago)

Quote:

fastfred said:
No fancy flow hoods or SABs.  Those can and do work, but they aren't as effective as good sterile technique with a bunsen burner.



How can you improve on 100% success?

I highly doubt your bunsen burner is any more effective than a SAB or Flow Hood that already delivers 100% success 99-100% of the time.

That is just silly.....effective is effective.

If they all give you great success they are all equal, none being more effective than the other.

Just use something to get the job done...if it works it works, but there is no need to exaggerate anything.

Extras: Filter Print Post Top
Invisiblefastfred
Old Hand
Male User Gallery

Folding@home Statistics
Registered: 05/17/04
Posts: 6,899
Loc: Dark side of the moon
Re: Sterilizing air of glovebox? [Re: PussyFart]
    #18879151 - 09/23/13 02:59 AM (10 years, 6 months ago)

Quote:

Notahacker420 said:
Quote:

fastfred said:
No fancy flow hoods or SABs.  Those can and do work, but they aren't as effective as good sterile technique with a bunsen burner.



How can you improve on 100% success?

Just use something to get the job done...if it works it works, but there is no need to exaggerate anything.




I'm not trying to exaggerate anything.  A bunsen in a draft-free room, combined with proper technique, is just fool proof.  It costs almost nothing and there's nothing to go wrong.  If the flame is lit then it's working 100%.

Poor flow hood design is standard, filters get clogged, punctured, and grown through, etc., etc..  The whole idea of whipping up the air with a big furnace blower has it's problems too.

I use a flowhood myself, but what it's useful for is larger size/scale stuff.  For most things it's just a fancy toy.

The problem is that people often use a flow hood as an alternative to good technique, and they end up skipping skills that are easy to learn and useful.  I don't see how you can ever learn good technique if your idea of doing micro is just to stick everything in a flow hood and assume everything is going to be sterile for you.


-FF

Extras: Filter Print Post Top
OfflineRogerRabbitM
Bans for Pleasure
Male User Gallery


Registered: 03/26/03
Posts: 42,214
Loc: Seattle
Last seen: 1 year, 1 month
Trusted Cultivator
OG Cultivator
Re: Sterilizing air of glovebox? [Re: fastfred]
    #18879431 - 09/23/13 07:04 AM (10 years, 6 months ago)

:facepalm:

This is just a total line of crap.  A burner will create turbulence and drafts just by running as it pulls air into the flame for combustion. I know of ZERO mycologists who use such nonsense.  All use flowhoods, and they're not just toys, they're tools which must be used correctly to be effective. 

To the original poster, you don't need to sanitize a glove or still air box.  The best technique is to spray it down inside with water.  This will attract and hold contaminants to the sides and bottom while you work.  For this technique, you want still air, not sterile surfaces since you won't be touching your sterile media to any part of the still air box.
RR


--------------------
Download Let's Grow Mushrooms



semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
Thomas Edison

Extras: Filter Print Post Top
InvisibleJuiceh
Dabbing All Day
 User Gallery


Registered: 09/25/12
Posts: 3,208
Re: Sterilizing air of glovebox? [Re: RogerRabbit]
    #18880046 - 09/23/13 10:43 AM (10 years, 6 months ago)

Bunsen Burner tek looks like it has some of the same problems as the Ceiling Fan tek, Stovetop Exhaust "flow hood" tek, and the Oven Door tek. With the added benefit of incinerating whatever gets caught in the updraft, which automatically makes it better than the Ceiling Fan tek and Oven Door tek but about on par with the stovetop exhaust tek, since it removes from the area whatever contams get caught in the updraft.

That drawing with the Bunsen burner clearly shows unclean air being pulled from the sides of the work space, right over your work. There's no way it can be claimed that air moving over the workspace is clean.

What happens when this unclean air is pulled over the surface of that petri dish when its opened?

This is what happens:



Look at the top right of the taller tower in Figure b, do you see how dirty air coming over the lip of your petri dish in this manner is a bad thing? The swirling air turbulence in that location will send contaminant spores right into your sterile media.

From what I can tell it looks to me that Bunsen burners were used back when we didn't know any better about how air currents really behave.


--------------------
Trade List

How I make 15 lbs of casing in a 941. How I line 12 shoeboxes with 1 39gal bag. How I Deep Fry Cubes!
RogerRabbit said:You need a bigger pressure cooker for this hobby. RR

Edited by Juiceh (09/23/13 10:44 AM)

Extras: Filter Print Post Top
Invisiblefastfred
Old Hand
Male User Gallery

Folding@home Statistics
Registered: 05/17/04
Posts: 6,899
Loc: Dark side of the moon
Re: Sterilizing air of glovebox? [Re: Juiceh]
    #18881879 - 09/23/13 07:20 PM (10 years, 6 months ago)

Quote:

RogerRabbit said:
This is just a total line of crap.  A burner will create turbulence and drafts just by running as it pulls air into the flame for combustion. I know of ZERO mycologists who use such nonsense.  All use flowhoods, and they're not just toys, they're tools which must be used correctly to be effective.




That's one of the biggest problems with the OMC.  Even experienced growers have learned what they know only from experience and other old hands.  You end up with a mix of wives' tales, superstition, dogma, ritual, and lore presented as the only way to be right.

It's pointless to argue this.  Anybody who's taken general microbiology lab already knows that bunsen burner work is how things have been done since the beginning of micro, and how the vast majority of it is still done today.



This is absolutely standard and how every microbiologist (including mycologists) is trained.  The fact that I'm constantly questioned on this every time I point this out goes to show that very few here have taken even a single lab course in micro.

Quote:

Juiceh said:
Bunsen Burner tek looks like it has some of the same problems as the Ceiling Fan tek, Stovetop Exhaust "flow hood" tek, and the Oven Door tek.




This isn't a "tek", this is the standard technique used by everyone who has ever had ANY lab training in microbiology.

Quote:

That drawing with the Bunsen burner clearly shows unclean air being pulled from the sides of the work space, right over your work. There's no way it can be claimed that air moving over the workspace is clean.

What happens when this unclean air is pulled over the surface of that petri dish when its opened?




The diagram doesn't show the larger pattern of air entrainment.  Just imagine the pattern being larger, and remember the hole is right at the bottom.  Even the lip of a petri dish will be at or higher than the hole.  Only a small amount of air is taken into the hole, the larger pattern is an updraft in the whole area around the bunsen.

Just imagine the arrows to the hole going upwards with a tiny amount of that upwards moving air sucked into the hole.

Quote:

From what I can tell it looks to me that Bunsen burners were used back when we didn't know any better about how air currents really behave.




No.  It was invented way back at the beginning of microbiology and is still used today because it works the same as it always has.  It worked great for Robert Bunsen in 1855, worked great when I took general micro lab, and work just as well today.

The basics of micro are... Always work in the most draft-free area possible, sterilize your work surface with 1:10 bleach and allow to dry, light your bunsen burner, flame your tool, keep close to your bunsen, and you're off and running.

Contams fall down in still air, always.  Contams cannot fall down into your cultures when you are operating in the updraft of your bunsen.

Why don't most labs use flow hoods?  Because they're expensive and unnecessary.  Spending $10k on a convenience doesn't make sense most of the time.

But the main reason most micro labs don't use them is that using a blower to blow air across your cultures and right into your face and onto your clothes is a bad idea.  You can't work with pathogenic or unknown cultures using a flow hood.  There's more expensive systems that contain the air, but they're VERY expensive to buy and maintain.


Man it gets boring arguing over first day topics of the first micro lab course.  You guys really need to open a book once in awhile!


-FF

Extras: Filter Print Post Top
OfflineRogerRabbitM
Bans for Pleasure
Male User Gallery


Registered: 03/26/03
Posts: 42,214
Loc: Seattle
Last seen: 1 year, 1 month
Trusted Cultivator
OG Cultivator
Re: Sterilizing air of glovebox? [Re: fastfred]
    #18884006 - 09/24/13 09:22 AM (10 years, 6 months ago)

:facepalm: Listen for the 100th time, since you've never grown mushrooms, but feel compelled to constantly post in advanced mycology.

I was in a microbiology lab yesterday to get several septic samples tested for the county conservation district.  They NEVER and I must repeat for you NEVER keep cultures longer than 48 hours.  This is why they can get away with using less than sterile technique, such as a Bunsen burner.  Contaminant spores simply do not have time to germinate and grow before the petri dish is discarded.

In mycology we keep the same cultures alive for 30 years or more.  In fact, my best Shiitake strain is over 30 years old, and my Oyster strain which is one of the best performing strains in the world is over 10 years old.  This requires 100% sterile procedure, which is why we wear surgical gowns, gloves, masks, hairnets, and run laminar flow hoods, usually in a positive pressure lab, charged by HEPA filtered intake.

Of course, you've had all this explained to you many times before. . .
RR


--------------------
Download Let's Grow Mushrooms



semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
Thomas Edison

Extras: Filter Print Post Top
Invisiblefastfred
Old Hand
Male User Gallery

Folding@home Statistics
Registered: 05/17/04
Posts: 6,899
Loc: Dark side of the moon
Re: Sterilizing air of glovebox? [Re: RogerRabbit]
    #18887671 - 09/25/13 02:07 AM (10 years, 6 months ago)

Point taken, again...

A microbiologist usually considers their cultures clean when they get good growth with no contamination after 3-7 days.  A mycologist might not get ANY growth for 3+ days, and can't consider it clean for weeks.

The principles are still the same.  Saying a SAB beats the updraft of a bunsen, or even a flowhood, is a matter of debate, to be settled by personal experience and results.

The fact is that you don't learn good technique working in a flow hood, and when shit goes wrong you're screwed with no clue.

There's no reason people without a flow hood shouldn't learn proper sterile technique.  And there's no reason that people WITH a flow hood shouldn't learn it, before relying on and betting the farm on it, first.


-FF

Extras: Filter Print Post Top
Offlineforrest
 User Gallery


Registered: 11/16/12
Posts: 1,011
Loc: The Netherlands
Last seen: 4 years, 8 months
Re: Sterilizing air of glovebox? [Re: fastfred]
    #18914419 - 10/01/13 02:28 AM (10 years, 5 months ago)

i'm in the first year of a biologyrelated study and asked second years how they dealt with bacterial cultures on agar, and they said they used a bunsenbrander and usually kept cultures for about a month.

(just some info)


--------------------
My Trade List

Extras: Filter Print Post Top
OfflineSkysTheLimit
A curious mind


Registered: 07/28/13
Posts: 168
Loc: Flag
Last seen: 8 years, 6 months
Re: Sterilizing air of glovebox? [Re: forrest]
    #18914498 - 10/01/13 03:14 AM (10 years, 5 months ago)

Quote:

forrest said:
i'm in the first year of a biologyrelated study and asked second years how they dealt with bacterial cultures on agar, and they said they used a bunsenbrander and usually kept cultures for about a month.

(just some info)




naahhh mycological work is FAR more susceptible to contaminants than bacteria colonies lol

Extras: Filter Print Post Top
OfflineRogerRabbitM
Bans for Pleasure
Male User Gallery


Registered: 03/26/03
Posts: 42,214
Loc: Seattle
Last seen: 1 year, 1 month
Trusted Cultivator
OG Cultivator
Re: Sterilizing air of glovebox? [Re: forrest]
    #18914666 - 10/01/13 05:26 AM (10 years, 5 months ago)

Bacteria cultures on agar are kept until identified and then discarded.  That's why they can use poor technique.  We keep cultures for many years, thus require 100% sterility.  None of this has anything to do with gloveboxes or sanitation for mycology work.

Stay on topic please.  A jet fighter pilot isn't going to take flight instruction from a kid who builds model airplanes.
RR


--------------------
Download Let's Grow Mushrooms



semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
Thomas Edison

Extras: Filter Print Post Top
Offlineforrest
 User Gallery


Registered: 11/16/12
Posts: 1,011
Loc: The Netherlands
Last seen: 4 years, 8 months
Re: Sterilizing air of glovebox? [Re: RogerRabbit]
    #18919795 - 10/02/13 07:14 AM (10 years, 5 months ago)

I wouldn't use a bunsenbrander for mycology and I don't advocate it. It was just a reaction to your statement about keeping cultures for only 48 hours.


--------------------
My Trade List

Extras: Filter Print Post Top
Offlinec-string
Brainless Layperson

Registered: 08/15/13
Posts: 6
Loc: The Void That Binds
Last seen: 10 years, 4 months
Re: Sterilizing air of glovebox? [Re: SkysTheLimit]
    #18930037 - 10/04/13 07:59 AM (10 years, 5 months ago)

Howdy, longtime lurker, first-time poster here. Once upon a time, I spent a decade working for DuPont in OEM automotive paint at a customer site. In the world of automotive paint, a lot of money and effort is spent to ensure that contaminants don't fall onto freshly painted surfaces and one strategy is to apply 'tacky coat' to spray booths and oven entrances. These are thin, sprayable coatings that remain slightly sticky in humid environments and thereby trap dirt (contaminant) particles.  Every so often you pull (or wash) the old coating off and reapply. I'd love to try them in my SAB but don't currently have a source. However, someone else may have access to something that would work along those lines. Since you can't totally prevent contaminants from getting in, the next best thing is to sequester them after they've arrived. No harm, no foul, as it were.

It's the same idea others have expressed in this thread about making sure that contaminants are precipitated out of the air and don't get stirred up, only more so.

This link gives you an idea of the sorts of materials I'm talking about.

Extras: Filter Print Post Top
Jump to top Pages: < Back | 1 | 2  [ show all ]

Shop: North Spore North Spore Mushroom Grow Kits & Cultivation Supplies   Left Coast Kratom Buy Kratom Extract   Original Sensible Seeds Bulk Cannabis Seeds   PhytoExtractum Kratom Powder for Sale   Mushroom-Hut Substrate Bags   Myyco.com Isolated Cubensis Liquid Culture For Sale


Similar ThreadsPosterViewsRepliesLast post
* sterile technique Killa_J 2,024 3 08/11/02 08:07 AM
by Anonymous
* Questions about a sterile glove box
( 1 2 all )
themills 4,825 28 08/14/04 12:19 AM
by phucknut68
* chemical sterilization by ethylene-oxide ? ragadinks 1,703 9 08/09/04 05:36 AM
by fastfred
* Bunsen burner?
( 1 2 3 all )
AsuraS 6,410 45 01/25/15 07:44 AM
by DrCubensis
* glovebox q and q K0r 1,178 7 03/15/02 11:37 AM
by BeppoMarx
* Inexpensive Glovebox.
( 1 2 all )
EffedS 4,241 22 05/26/03 09:23 PM
by Effed
* Glovebox critiscism/questions? socratesmind 1,043 2 10/07/02 09:23 PM
by Green_Velvet
* Sterility dioze1 2,116 14 09/03/01 06:57 PM
by fuzzysquirelnuts

Extra information
You cannot start new topics / You cannot reply to topics
HTML is disabled / BBCode is enabled
Moderator: RogerRabbit, Pastywhyte, bodhisatta
7,867 topic views. 0 members, 3 guests and 4 web crawlers are browsing this forum.
[ Show Images Only | Sort by Score | Print Topic ]
Search this thread:

Copyright 1997-2024 Mind Media. Some rights reserved.

Generated in 0.025 seconds spending 0.005 seconds on 13 queries.