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RNA in native gel


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3 replies to this topic

#1 hianghao

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Posted 01 March 2011 - 12:31 AM

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

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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.

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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?

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


#2 hianghao

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Posted 02 March 2011 - 01:10 AM

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


#3 merlav

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Posted 03 March 2011 - 07:10 AM

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
Science without religion is lame, religion without science is blind.
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I am among those who think that science has great beauty. A scientist in his laboratory is not only a technician: he is also a child placed before natural phenomena which impress him like a fairy tale.
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#4 hianghao

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Posted 03 March 2011 - 07:55 AM

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.





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