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Getting sterile Agar from PC to my Glovebox


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#1 Guest_SnakeEyes_*

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Posted 18 July 2006 - 09:40 AM

I just want to verify my procedure for agar based petris. I pc my malt extract, take directly out and put in the glove box, my prob is finding something to pc it in that will fit in my glove box. I would say like pour into a small pitcher, but that would need to be sterlized before I work. Getting the hot agar to my glove box, uncontaminated is my main goal for this post. Please "Help a brotha out"

#2 siam_jim

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Posted 18 July 2006 - 09:52 AM

i use are soda bottles fill them 1/3 and plug it with cloth and top off with foil and rubber band it. works great! after pc'ing just take it to the GB and open it...simple no contams

siam

#3 Guest_SnakeEyes_*

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Posted 18 July 2006 - 10:02 AM

great thinking siam. i wasnt sure what materials could withdstand PC, although its only 15 mins. Thanks man.

#4 siam_jim

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Posted 18 July 2006 - 10:21 AM

also go 20 at lease, i do mine for 25-30,

don't open the bottle till its cool enough to pour as heat will rise and cause a suction to be drawn into the bottle along with possible contam. i usually leave it in the pc till ready tho.


siam

#5 Guest_SnakeEyes_*

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Posted 18 July 2006 - 12:20 PM

Just to clear things up, were talkin plastic bottles right?

#6 siam_jim

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Posted 18 July 2006 - 12:29 PM

oh NO! use glass soda bottles like the one in the pic

siam

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#7 Guest_SnakeEyes_*

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Posted 18 July 2006 - 01:29 PM

Damn those are hard to find anymore. I could substitute like a glass vinegar bottle or something similar Im sure. Thanks siam.

#8 sandman

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Posted 18 July 2006 - 06:59 PM

or just a quart jar with pollyfill lid, or whatever size jars you have. Kinda harder to pour however.

#9 python

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Posted 18 July 2006 - 07:53 PM

best thing to use is a "florence flask"---do an internet search or try mycosupply

or one can use a soda bottle like above just fine----or an erlenmyer flask....



simply stuff cotton in the top of the flask/bottle and cover the top with foil or even a double foil......once PC'd take to your glove box or flowhood.....

#10 Guest_SnakeEyes_*

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Posted 18 July 2006 - 08:30 PM

yea its heath lol :flamer: how'd you know?

#11 python

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Posted 18 July 2006 - 08:32 PM

:reb:

your question/point of interest and they way you type, told me!

#12 Guest_SnakeEyes_*

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Posted 18 July 2006 - 08:53 PM

Yea tru that. Anyways Mycosupply.com has all I need it's just, the site is wessed up, I cant click on anything. Comes up "Internal Server Error". I left em a msg, and Im callin back tomarrow so well see bout that one. But thanks for referal. -SnakeEyes

#13 BuckarooBanzai

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Posted 18 July 2006 - 09:00 PM

My FOAF uses a quart jar with two silicone injector ports. The jar is filled no more than 2/3 with agar mix and PCed for 25 min @ 15 psi. When cool enough to handle, the jar is moved to the flow hood (or in your case, glove box).

An airport syringe is flamed and then injected through one port to release the vacuum formed in the PC. The airport is left in place (so a vacuum will not reform) and a 60mL syringe (sterile) is injected through the second port. The 60mL syringe is filled with hot agar and then withdrawn from the jar.

Plates are then filled (in front of the flow hood) by opening them only enough to allow the 60mL's needle in. Plates are NOT opened all the way. The 60mL syringe's plunger is then depressed enough to put about 9mL of agar into the plate (enough to just cover the bottom of the plate).

The plates are stacked immediately after filling and set in front of the flow hood until cool. Cooling them in a stack will prevent all but the top one from forming a ton of condensation on it's lid.

As soon as the plates are cool, they are sealed with a double layer of Parafilm. The filmed plates are then double bagged in freezer grade Ziplocs, dated with a Sharpie and kept in the fridge until needed.

Plates are all used within 2 months or tossed.

#14 Guest_SnakeEyes_*

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Posted 18 July 2006 - 10:18 PM

Thanks for your input Buckaroo, so you say flow hood, but ill be letting mine cool in the glovebox, correct? And your Saying I have to wait till they cool the cover with parafilm(btw wuts the difference between parafilm and clingwrap? tiny airation hols?) also would mycosupply carry parafilm or can i get it locally? Thanks- Snake

#15 python

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Posted 18 July 2006 - 11:02 PM

parafilm is at mycosupply

it is a labwork product that can stretch, seal, and on top of that; it is waterproof

#16 Guest_SnakeEyes_*

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Posted 18 July 2006 - 11:06 PM

Thanks Python.

#17 BuckarooBanzai

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Posted 19 July 2006 - 08:15 AM

Dig – cool ‘em in the glove box. You could wrap them right after pouring, but since the agar will still be liquid, you’ll have to be careful or it will slosh/spill. Petri dishes do NOT make a good seal (Petri’s make no seal at all, really).

You could use cling wrap or masking tape or any number of other things, but Parafilm works MUCH better. Don’t be put off by the price, one roll of it will last forever. If you just want a small amount to play with, go check out eBay. Lots of folks sell it in short strips (it is used by model builders to mask areas as they paint).

If you are having trouble with the Mycosupply site, you might consider trying Sporeworks.com

It’s just opinion, but I’ve always found Mycosupply just a little sketchy for some reason that I can’t quite put my finger on.

#18 Hippie3

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Posted 19 July 2006 - 09:05 AM

My FOAF uses a quart jar with two silicone injector ports. The jar is filled no more than 2/3 with agar mix and PCed for 25 min @ 15 psi. When cool enough to handle, the jar is moved to the flow hood (or in your case, glove box).

An airport syringe is flamed and then injected through one port to release the vacuum formed in the PC. The airport is left in place (so a vacuum will not reform) and a 60mL syringe (sterile) is injected through the second port. The 60mL syringe is filled with hot agar and then withdrawn from the jar.

Plates are then filled (in front of the flow hood) by opening them only enough to allow the 60mL's needle in. Plates are NOT opened all the way. The 60mL syringe's plunger is then depressed enough to put about 9mL of agar into the plate (enough to just cover the bottom of the plate).

The plates are stacked immediately after filling and set in front of the flow hood until cool. Cooling them in a stack will prevent all but the top one from forming a ton of condensation on it's lid.

As soon as the plates are cool, they are sealed with a double layer of Parafilm. The filmed plates are then double bagged in freezer grade Ziplocs, dated with a Sharpie and kept in the fridge until needed.

Plates are all used within 2 months or tossed.



:cool:




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