15 replies to this topic

[tamarine]

Hi,
I am newbie in PAGE.

I was wondering if somebody could help me.
I use SE 600- Amersham Gel caster for my PAGE. But, why every time i cast my gel, it always torn once i remove the glasses. the gel sticks to side by side with the glasses tightly. Do i have to use the repellent chemicals? if that so, would you mind to give me the brand. Are there any stages, i have to mind before pouring my gel to the caster?

I am very desperate. Really need the solving problem.. Thank you.

[bob1]

You need to clean the glass plates and any spacers before use with 70% ethanol and make sure that you remove any dried acrylamide on the plates during this process.  It is easiest to clean all these soon after taking the gel out - wash under running water such that the water forms a laminar flow over the plate.  This will allow you to see any residues on the plates, which can then be rubbed off with your fingers.

You can also coat one plate with a silicon repellent.  Rain-x is the stuff we have here, you can get it from car service shops etc.

[tamarine]

bob1, on 29 June 2011 - 12:55 AM, said:

You need to clean the glass plates and any spacers before use with 70% ethanol and make sure that you remove any dried acrylamide on the plates during this process.  It is easiest to clean all these soon after taking the gel out - wash under running water such that the water forms a laminar flow over the plate.  This will allow you to see any residues on the plates, which can then be rubbed off with your fingers.

You can also coat one plate with a silicon repellent.  Rain-x is the stuff we have here, you can get it from car service shops etc.

Hi Bob1, thank you for the advice.
Rain-x is good to considered. Soon, i am going to purchase it. How long can i wait the dry Rain-x before i pour my gel?
And what about the other plate? Do i have to stick it? Cause, i have to transfer the gel for the visualization stage. Is it ok to treat both glasses?

Your advice is going to help me. Thank you...

[bob1]

Rain-X is the sort where you rub it on using a cloth, and then a couple of minutes later polish it off.  You should only coat one plate so that the gel doesn't accidentally slide off the other plate while you are trying to manipulate it.

[tamarine]

bob1, on 02 July 2011 - 01:54 PM, said:

Rain-X is the sort where you rub it on using a cloth, and then a couple of minutes later polish it off.  You should only coat one plate so that the gel doesn't accidentally slide off the other plate while you are trying to manipulate it.

Ok. i am really grateful of your kindness. i can't wait any longer to practice it.
btw, have you ever work with the flatbed? it provided by Multiphor II Electrophoresis system (GE). Currently, i am working with this instrument and it quite got my annoying. once again, i ask you for a favor. if you don't mind, give me a little experience related to this. Thank you...

[bob1]

Sorry, never used the flatbed system.

[tamarine]

bob1, on 04 July 2011 - 03:02 PM, said:

Sorry, never used the flatbed system.

Ok, anyway i am really grateful for your sharing.

[Inmost sun]

coating can be done with diluted silanes or ready-to-use products like Sigmacoat

[tamarine]

Inmost sun, on 08 July 2011 - 12:10 PM, said:

coating can be done with diluted silanes or ready-to-use products like Sigmacoat

Hi,
Currently, I was utilizing the Sigmacoat. But, i don't know why, it seems can't work. The gel is still bind at side by side to the glasses. Do i miss something essential due to preparation? How do you use the chemical before pouring the gel to its caster?

Thank you for the response,

[Inmost sun]

I think Sigmacoat is a silane diluted in organic phase; it works for glass but not alox plates (Hoefer); plates must be cleaned, then coated by rubbing a portion with cloth, shortly rinsed with distilled water and dried;

if Sigmacoat does not work in your case it seems to be too diluted for your purpose; you may repeat coating twice or more;

or buy pure silane (Sigma)and dilute with CCl4

[phage434]

Sigmacoat also has a limited lifetime, so old samples may not work well, or at all.  Clean glass is an absolute requirement.  Scrubbing with alconox or a similar detergent is a good start, but you need to get the glass clean enough so that water sheets and flows over the surface with no beading up.

[tamarine]

Inmost sun, on 09 July 2011 - 12:08 PM, said:

I think Sigmacoat is a silane diluted in organic phase; it works for glass but not alox plates (Hoefer); plates must be cleaned, then coated by rubbing a portion with cloth, shortly rinsed with distilled water and dried;

if Sigmacoat does not work in your case it seems to be too diluted for your purpose; you may repeat coating twice or more;

or buy pure silane (Sigma)and dilute with CCl4

Hi, Inmost sun.
Usually, i was practicing the rub with Kimwipes. is it not good enough for rubbing? and what am i supposed to do? should i purchase the new chemical or substitute the glass?

Pardon me, what kind of CCl4 is? i don't ever hear it?
thank you for the guidance.

[tamarine]

phage434, on 09 July 2011 - 12:16 PM, said:

Sigmacoat also has a limited lifetime, so old samples may not work well, or at all.  Clean glass is an absolute requirement.  Scrubbing with alconox or a similar detergent is a good start, but you need to get the glass clean enough so that water sheets and flows over the surface with no beading up.

Hi, phage434.
Thank you for the response. Frankly, your hypothesis is good to think. i am gonna check the expired date of my chemical,soon. Would you share me about alconox? is it Extrant (Merck brand)? actually, my lab posses it. and it is a kind of detergent. but, still i cant find the answer conducting these mess.

Your kindness is highly appreciated.

[mdfenko]

tamarine, on 16 July 2011 - 05:02 AM, said:

Pardon me, what kind of CCl4 is? i don't ever hear it?

ccl4 is carbon tetrachloride, a solvent.

we used rain-x for our sequencing gels.

Edited by mdfenko, 18 July 2011 - 11:52 AM.

[tamarine]

Thank you mdfenko!!

I gotta do this sooner! Thank you. Thank you.