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Northern blot with a RNA probe labeled with a fluorophore

northern blot RNA probe fluorescent RNA probe

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#1 hudgie



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Posted 11 May 2018 - 07:12 AM

I have some fluorophore labeled RNA probes that I want to try in a Northern Blot analysis using Qiagen RNeasy kit purified RNA.

I want to image the blot on the LiCor Odyssey imager/scanner. They recommend using a Northern Max kit with DIG labeling of the RNA probe prep by RT-PCR but I already have fluorescent labeled custom probes that I don't want to waste.

Every northern blot protocol I see for RNA is for DIG-labeled probes. Does any have any "protocol suggestions" for me to use these expensively made custom fluorophore RNA probes??

#2 bob1


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Posted 11 May 2018 - 08:15 AM

Basically if you have the probes already, just use those in the place of the probes in the DIG protocol. There might be some subtle differences in binding of a RNA probe vs DIG-labeled DNA, so you might have to play around with the conditions a little. If I recall correctly, there are some conditions for RNA probes somewhere in the DIG handbook.

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