RNA in native gel - (Mar/01/2011 )
Hi, I ran 2 RNA native gel to qualify my RNA for qpcr. 1h on 1% gel. Gel photos were attached.
http://img826.imageshack.us/img826/9733/hhrna1.png
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.
http://img808.imageshack.us/img808/2166/rna2.png
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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?
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?
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.com/techlib/append/supp/rna_gel.html
merlav on Thu Mar 3 15:10:02 2011 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.com/techlib/append/supp/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...