3 replies to this topic

[hianghao]

Hi, I ran 2 RNA native gel to qualify my RNA for qpcr. 1h on 1% gel. Gel photos were attached.


Are the last 2 lanes degraded RNA? What bout the 1st four lane? Is my separation time too short? Samples are incubated with formamide+formaldehyde at 65C for 15 minutes before loading.



Uploaded with ImageShack.us
This gel is a bit weird because the bands are of length 2000bp and 1000bp... Is it because i didn't add and incubate the RNA with formamide+formaldehyde?

Edited by hianghao, 01 March 2011 - 03:25 AM.

[hianghao]

Somehow i cant find the "edit" button...
This is my new gel using the same samples and the results are more or less the same. Which samples i use for qPCR?

Edited by hianghao, 02 March 2011 - 01:11 AM.

[merlav]

Run a Denaturing gel instead of a native gel, because in native gel the RNA has the secondary structures and afect migration. check this link http://www.ambion.co...pp/rna_gel.html

[hianghao]

merlav, on 03 March 2011 - 07:10 AM, said:

Run a Denaturing gel instead of a native gel, because in native gel the RNA has the secondary structures and afect migration. check this link http://www.ambion.co...pp/rna_gel.html

I hope i can but i dont have MOPS... The loading dye contains formamide so the supplier says that it can be run on native gel though...

Edited by hianghao, 03 March 2011 - 08:04 AM.