Abnormal Taqman qPCR multicomponent plot caused by Taqman probe degradation?

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Does anyone know HRM channel in a 2 plex QIAGEN Rotor- Gene Q qPCR can be used as a separate channel?

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How to address this issue in the analysis of Real-Time PCR results?

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No title

Does anyone have the experience of using Taq Man probes in the QIAGEN Rotar- Gene qPCR machine?

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Is CD44 degraded in lysosome together with hyaluronan after CD44 mediated endocytosis of hyaluronic acid?

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MiRNA qpcr problem?

Dear All, mirna primer showing some problem in the melting curve? any idea why? As attached is the melting curve. The forward sequence is obtained from miRBase and reverse primer is universal.

28 February 2021 5,008 4 View

Kevin Squires

My first guess would be there is too high a concentration of template DNA in the "abnormal" reactions. Did you standardize your template concentrations using a nanodrop?

Jia-Wang Wang

Yes, we did Nanodrop and the concentrations should be the same.

Christian D Ahrberg

To me it also looks like a too high template concentration. Have you tried doing a dilution of your sample. Looking at your curves i would assume you should try diluting by a factor of 100 to 1000.

I would be very happy if it is like you said but doubt it because we were detecting plasma miRNAs which are usually low