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Here's my GB....anticipating all that follows..[ glovebox merged ]


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#1 Old_Recordings

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Posted 12 October 2005 - 11:25 PM

Recently the idea of a glovebox came to mind, with that a couple of questions followed. Using plastic "Big Gulp" cups cut in half for glovebox holes makes it very easy to attach rubber gloves to with rubber bands to hold 'em in place. Would sealing these cups with caulk silicone hold them in place and resistant to contams? Also, the lid of the Rubber Maid box is not air tight, browsing the archives and finding out that "seran wrap" should be applied to the edges of the box for a tighter seal. I'm unclear on how to apply the wrap to the edges of the box. If anyone know anything about this procedure please reply. Thanks. Peace.

#2 Guest_golly_*

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Posted 13 October 2005 - 06:33 AM

Using plastic cups as a mounting device for gloves can work...i found 41/2 inch diameter plastic flower pots that are a bit more ridged than most cups.
The gloves seem to cling well without any reinforcement..
Make sure u can slip your hand in and out without too much resistance...
The best glue i have found for bonding the glove assembly to the box is "Goop" -make sure to roughen both surfaces with sandpaper to increase the strength of the joint...As for sealing the lid - it's hard to know without seeing the box but a combination gasket +tape or a weight ..has to be easy to get in and out of...gluk...

#3 sandman

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Posted 13 October 2005 - 01:01 PM

indeed glueing them with something would make it stronger, and also cut down on accidental airflow. Something that DOESNT work very well is hot glue. It will hold for a second or 3, but with frequent use it will lose its bond. Strongest possible bond will be epoxy resin, or a 2 part epoxy glue. Listen to golly about the sanding of the area to be joined, its great advice and makes all the difference on the strength of the joint. You can mix the epoxy glue with pulverized limestone (something a grower might have) or better, wood flour (very very fine sawdust) or even regular flour would work and make what is known as a filet on the joint to achieve ultimate strength. Basicaly you add a bead of the peanut butter consistancy filet material to glued joint (when wet) and use a scraper with a rounded corner to make a nice round concave joint (imagine a U flush against a right angle). I guarantee you will never rip the armholes out in the middle of sterile work like I have done so many times with the sorry ass hot glue. Maybe the Goop wood work as well, just srape the joint with something U shaped to get convex shape of the joint.

My box is a rubermaide and the top is indeed not very airtight. I dont mess with no saran wrap except the window cut in the lid in the middle is saran wrap duct taped. I have heard of people putting down some weather stripping to help the seal. I dont worry about it myself. Maybe I should.

#4 Hippie3

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Posted 14 October 2005 - 11:59 AM

Using plastic "Big Gulp" cups cut in half for glovebox holes makes it very easy to attach rubber gloves to with rubber bands to hold 'em in place. Would sealing these cups with caulk silicone hold them in place and resistant to contams?


i did just exactly that with one of my earlier 'boxes,
worked well enough.

#5 Old_Recordings

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Posted 14 October 2005 - 01:59 PM

Thanks for the feedback fellas. Very useful info. Peace.

#6 mr_dimple

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Posted 17 October 2005 - 11:44 AM

OK, so my pet rabbit has got a 54 quart plastic container that he’s turned into a glovebox – no positive pressure, just a good seal all around - a sheet of fiberglass on top for good visibility, well sealed, and rubber gloves attached to glove holes, also sealed.

It’s my rabbit’s first time using a glovebox and after shooting 6 BRF jars yesterday, he finds he has some questions about how to best sanitize his glovebox and what workflow to use to successfully inoculate jars.

Should he be spraying the inside of the glovebox with Lysol or bleach? Yesterday, he used a store-bought Clorox-Clean-Up spray bottle, but he’s heard people in the archives talk about diluted bleach, and wonders what on earth that is.

Should he be spraying his glovebox before he puts jars and syringes inside or after? And should he be worried the fumes will do unmentionable things to his spores?

Should he be wiping the tops of his jar lids with bleach after removing the foil and before shooting them up? Yesterday, my rabbit used a lighter to sterilize the syringe needle between each jar, should he continue doing that next time?

(Also, my rabbit had a hard time controlling the flow of liquid from his syringes and is pretty sure a couple of jars got over 2ml of sweet spore water, not what he had in mind.)

My rabbit would greatly appreciate a step by step workflow from any of you who are using or have used a similar glovebox set-up.

Just fyi, the first time my rabbit tried his hand at microscopy, he simply used a clean bathroom for spore injection so he’s finding the glovebox extremely cumbersome, but since he spent a few hours building it, he is committed to learning how to use it well :-]

:gah:

#7 laborer

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Posted 17 October 2005 - 12:00 PM

Some use bleach water, I use lysol myself. I wipe everything down with lysol that I put into the gb. Then i spray some lysol into the gb. Havent had any contams doing g2g transfers. Only downside to lysol, its is extremely flammable so you dont want to sterilize a needle with a flame in a gb. I just fill a shot glass with rubbing alchohol and put that in the gb and dip my needles in it....

#8 Guest_dial8_*

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Posted 17 October 2005 - 01:18 PM

The nature of a glovebox is to be cumbersome unless you have built a really good one. I usually reccoment thinking it through really well so you can build the most comfortable. I usually sanitize my jars and all of my untensils. I sanitize the gb then I place all the stuff inside. Then I spray the inside really weell again and immediately place the lid on. Use a 10:1 water bleach solution or some lysol. IF YOU USE LYSOL GIVE IT AMPLE TIME TO SETTLE IF YOU WILL BE USING A FLAME INSIDE THE GB. If you don't it will go KABLOOM!
I would reccommend waiting at least 15 minutes before starting.

#9 Guest_freakachino_*

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Posted 17 October 2005 - 02:29 PM

I would always just wipe the inside down with an iso alky sponge, then wipe everything going in it down with alky, close it up. Then I go to work. It is meant to be still air so all work should be clean. Also, I remove all foil thats been in my pc before I put anything in my gb. Wiping everything down seems to work for me and no waiting period or explosion possibilities with the lysol. The alky evaporates pretty quickly. I also use 91% iso in my gb for this reason.

They are a pain to work in but I found after the first few tries I was used to it enough and happy that I could do sterile work without worries of contams or sterilizing an open area. The most cumbersome is the gloves and where your arms enter. If you can find a way to make that comfy, your rabbit will be happy :)

#10 Hippie3

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Posted 17 October 2005 - 09:47 PM

what you need depends on what you are doing.
pf style jars can safely be inoculated in open air,
no glovebox needed.

#11 mr_dimple

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Posted 18 October 2005 - 08:55 AM

Well, my rabbit is doing some pf style half-pints but also some liquid cultures (Karo) and is planning on trying grain to use as spawn for casings (worm poo). His first attempt at pf style cakes, a few years back, was not a complete failure - two flushes on 12 cakes (10 survived), so now he wants to try other methods, thus the glovebox.

My rabbit was considering wiping down jar lids with alcohol before injection, but wasn't at all sure that would do the job, given how quickly the 91% iso evaporates, but he will take freakachino's advice and try it next time.

Thanks for all the other advice, too, though my rabbit is always happy to get even more pointers so he can become a Zen Master of Cleanliness!

:bow:

#12 Guest_Peter Cottontail_*

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Posted 18 October 2005 - 09:09 AM

Use 70% alcohol to wipe down jar lids and your gloves. It works better.
RR

#13 Hippie3

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Posted 18 October 2005 - 09:12 AM

yeah,
for not-so-obvious reasons
the 70% seems more effective than higher concentrations.
less explosive/flammable, too, although still dangerous.

#14 mr_dimple

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Posted 18 October 2005 - 09:23 AM

So, at the risk of making my rabbit sound like a retard - while inside the gb - take off foil, wipe lid, inject, put foil back on?

Or should I not even bother putting foil back on? My incubator is running at ~82F so I will get some condensation and was thinking the foil will prevent the condensation from leaking into the jars.

:eusa_shif

#15 Hippie3

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Posted 18 October 2005 - 09:25 AM

i like to put foil back on, keeps dust off the lids.
btw
i'd lower the heat to 80,
glass jars trap heat so it's hotter inside.

#16 mr_dimple

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Posted 18 October 2005 - 09:43 AM

Thanks hippie, will lower the temp. The incubator is still a bit of a work in progress, the 100W fish tank heater I have in there is taking too much space, so I'm replacing it with a smalller 50W today.

One (hopefully) last question - what's the best way of sterilizing the syringe needle?

laborer (up above) suggested dipping it into alc, but I just got some really long (3') needles for my Karo jars and there's no way I can figure out a way to dip them, since they're so damn long. Should I be sterilizing the needle between jars to prevent cross-contamination?

#17 viewer6922

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Posted 26 February 2006 - 06:16 PM

I am new to this! Very excited about growing the little fellars....I built the GB with stuff laying around my palce, I did a BiB(box in box), measured and cut even more cardboard, cover that with aluminum foil, sealed the seams with clear duct tape,tapercut 'robo arms' from flexie dryer duct,then siliconed those in, and a piece of PlexiGlass duct taped on top,....measures a whopping 20"x17"x15"....so very much looking for my first crop by my birthday! April the first, spores are on the way, pressure cooker tested, jars are bought,which I bougth 24 to start, thinking about buying 12 more, and tommorrow, buying a fish tank heater and doing TiT for warmth, then I will build my fruiting chamber! I love the shroom, really looking forward to all! Thnks for all the help folks!



#18 Hippie3

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Posted 26 February 2006 - 07:01 PM

a few pix of yer box would be nice...

#19 doobydoobydoo

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Posted 26 February 2006 - 07:34 PM

even a diagram :)

#20 viewer6922

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Posted 26 February 2006 - 08:46 PM

sorry about the up loads....took me time to figure the stuff....soooo... what do yall think ? I will have 'rubbermaid' gloves on, 'kerchief over mouth and nose, antibacterial shower, hairnet, AND ! knocking up 36 jars!
Question! How often to I sterilize the needle per syringe? If I am changing syringes...do I only sterilize per syringe?
If I get a great batch, ie, alot of viable jars, what do you think of casing a few, and what TEK do you recommend for casing?:flamer:

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