Hello,
We are going to have a qPCR validation experiments from 108 sera (27 NSCLC Pre-Op, 27 NSCLC Post-Op, 27 High Risk Cohort, 27 Health) for 5 miRNAs.
We are planning to use spiked-in miRNAs as control in miRNA isolation, cDNA generation and qPCR steps. However, there is a need for an endogenous reference miRNA for serum to be used in data normalization.
Does anyone know any rescent articles mentioning any such refence miRNA in serum to be used in this purpose? I was unable to find.
Thanks in advance,
-Güven
Abbas Güven Akçay
Here I have attached some article used synthetic cel-miR-39 as a endogenous control.
Sincerely
Prabu
Thanks for your response. But I am confused. How can synthetic C. elegans miRNA can be used as an "endogenous" control. Did you mean it is used as an exogenous reference to normalize the data?
Abbas Güven Akçay
Not only exogenous control and some other types of normalization methods has been used in earlier publications such as
1)Endogenous control
2)geNorm
3)Normfinder
4)Global mean.
So, you can go ahead with anyone of above this and which one suitable to your hypothesis.
Sincerely
Prabu
let me share this document, which is a table of normalization methods from several similar studies, I obtained from the following paper:
doi: 10.18632/oncotarget.21739 PMID: 29212284
From your shared document 2 studies used exogenous control for normalization.
Foss et al. [27]
Li et al. [43]
Best wishes
prabu
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