hi

I am attaching some sourcez you may find informations. Regarding the optimal temperature now 45-55 approximately. Some more experients might add onto that .

Regards

Kat.

http://cmdr.ubc.ca/bobh/method/acid-urea-gel-electrophoresis/

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3329804/

https://molbio.mgh.harvard.edu/szostakweb/protocols/denaturepage/index.html

http://www.protocol-online.org/prot/Protocols/Denaturing-Urea-Polyacrylamide-Gel-Electrophoresis--PAGE--Based-Microsatellite-Analysis-3466.html

https://experiments.springernature.com/articles/10.1007/978-1-62703-565-1_11

30-40 volts works well

I dont have any Idea

Sorry

We use 10-15W pre-run and run. We don't do it in cold room. Only native gels we run in cold room.

Thank you Katerina Kouzari , Vasudeva Murthy and Jana Schmitzova for your reply.

You are welcome , keep us posted how it will go

Best of luck !

Thank you for all your help. The gel turned out to alright.

Katerina Kouzari : urea starts to decompose at temperatures above 50C. Urea decomposes to isocyanic acid which, in solution at least, causes irreversible carbamylation. I am not sure though about the likelihood of this occurring during a PAGE. But I would reduce the temperature.

Dear Achim

I ve also know that , but i ve check theres differences regarding the optimal temperature which depends upon the gel concetration, the current if its stable or not the pre run if the intesitu was run on constant 15mA for an hour for istance and last were we most get confused the voltage or watts will be used on separation. So far three different prptocols noted by websites do not indicate on the exact same temperatures (besides those were for incubation) or intensity. As for instance the intensity would have been high means it would also cause denaturation. Knowing what you say the most safe factor to induce you are absolutely temperature, but current also does. about PAGE i am attaching a troubleshooting guide i found

it is better to do your run at cold temperature to get good resolution and clear bands without distortion. The used voltage is around 40.

Good luck.