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denature RNA and miRNA Q-PCR


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#1 lymphomaW

lymphomaW

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Posted 04 February 2009 - 08:26 AM

I prepared my total RNA using mirVANA miRNA isolation kit. Then I heated the RNA @58 degrees for 10min and store the sample in -80 freezer. Now i'm going to perform a Taqman miRNA Q-PCR. When i read their protocol, they suggested not to denature the RNA because it might reduce the yield of cDNA for some miRNA target. Has anyone used denatured RNA for Taqman miRNA assay? Would the denaturation step be a big problem?

Thanks

lymphomaw

#2 microlight

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Posted 04 February 2009 - 10:49 AM

I remembered the protocol saying do not denature RT-primers for miRNA when prepare cDNA.




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