How does the PCR amplification of the Endogenous control targets differ from the amplification of the microRNA target?
Both the miRCURY LNA™ microRNA assays and the Endogenous control assays are based on a gene-specific RT primer for first strand cDNA synthesis. PCR amplification is then performed using one microRNA specific primer and the Universal primer, whereas the Endogenous control assay uses two gene-specific primers. This difference in assay design does not affect the usefulness of the Endogenous controls as normalizers for microRNA expression levels. Please see question 25 for details on how to correct for amplification efficiency.
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