Burning of IPG-strips

[holger_loerchner]

Dear colleagues,

my IPG-strips are burning during focussing over the last few weeks. In particular, the region between both electrodes is predomnantly burned/ vanished. I already checked or rather changed strips, rehydration solution, samples and so on but nothing really changed. Even classical advices from troubleshooting guides don´t help.
Does anyone got an idea ???


Best regards Holger

[admin]

Hello there Holger,
(a) Use only "THICK" mineral oil (US Biochemicals Cat.# US71600. This can also be purchased directly from Amersham Pharmacia. Amersham Pharmacia Cat. # US71600-1l). "THIN" mineral oil easily moves from well to well and may result in drying/burning of IPG strips during IEF.

(b) Ampholytes exceeding 0.5% may cause IPG strip to burn during IEF due to excess salt concentration.

Maybe this or other problems:

It may seem you either have a bad batch or may not be fully covering the strips with buffer.

cheers!

[proatpcr]

Hey there,
my IPG strips also started burning in the middle of electrophoresis.

When I used heavy mineral oil this never occurred though.

Are you using light or heavy mineral oil?

Light mineral oils can't withstand the hot temperatures during IEF and because of their light density they can easily flow from well to well of IEF focusing tray.

Please post back here maybe I will get this problem again in the future
good luck

[danfive]

Here is another mineral oil, I've never had issues with it Bio-Rad Mineral Oil Biotechnology Grade (cat# 163-2129).