what is the meaning of de -gas the acrylamide gel mix? - (Jun/28/2006 )

hi folks
i read in one of the posts here in the forum :
dont forget to de-gas your acrylamide gel mix under vaccum before adding TEMED and APS for 30 min
what is the meaning of de -gas the acrylamide gel mix???
why?
i dont do it really ???
thankx in advance

-spanishflower-

QUOTE (spanishflower @ Jun 28 2006, 10:41 PM)

De-gas removes the oxygen in the acrylamide gel mix as oxygen interfere with the polymerization of the gel.
I don't degas the gel mix......it just takes a bit more time to polymerize the gel.

I hope this may help.

-Minnie Mouse-

thankx minnie so much
but after i put my running gel into the glass slides i pour sometimes water and sometimes isopropanol around 1 ml on top to prevent air contact and usually it takes 15 min to polymeraise ( 12% gel) is this ok?
another question do have any usuful links for more information about SDS PAGE , i mean to understand the science behind every step??
thankx alot

-spanishflower-

QUOTE (spanishflower @ Jun 28 2006, 11:32 PM)

It is OK.

-Minnie Mouse-

The Sambrook and the Lane books say to use water, but I prefere to use isopropanol because I feel more confortable knowing that it will not mix with my gel. After that one has to rinse a couple of times.
This is a link I gave to one of my students: it's very basic, probably too easy for you but you can give it a look

http://www.davidson.edu/academic/biology/c...GE/SDSPAGE.html

-dnafactory-

THANKX REALLY.

-spanishflower-

Spanish flower, if not de-gassing woks, great. The issue is that wee tiny bubbles in the gel can interfere with the flow of proteins and give you a distorted or even erroneous gel. To be honest, I think a lot of us just settle for a good looking gel (i.e. no visible bubbles).

In HPLC we have to de-gas so that the pump action is smooth and no air-locks occur in the pipes or column.

-paraboxa-