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miRNA rtqPCR

grrompala
in Medical Science

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Hi,

 

I'm using the Qiagen miscript rtqPCR kit for small RNA quantification. I did a gel purification for RNA bands >40bp and those <40bp. When I then performed miscript with miRNA specific primers, I still had a strong signal ~24 ct for the >40bp gel fraction. However, melt curves and pcr products looked identical between >40bp and <40bp. Could it just be my gel run wasn't efficient in separating the bands? Seems hard to believe. Any insight would be helpful, thanks!

Edited by grrompala

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