Simon Baker 1,
1Bioline Reagents Limited, United Kingdom
For some time now, Bioline has offered an extensive list of indi- vidual human miRNAs, using the proprietary algorithm developed by MiRXES to maximize miRNA detection sensitivity, while mini- mizing non-specific interactions. The versatility of the assay design system has now allowed us to extend the range to include all the 27,000 miRNA listed on miRBase, providing plant and animal miRNA assays. The resulting real-time PCR assays enable detec- tion of extremely low levels of miRNA with high specificity using a SYBR® Green detection chemistry, allowing the discrimination between closely related miRNA sequences. All EPIK miRNA Select Assays have been validated using synthetic miRNA templates and human assays have also been validated against total RNA. Typically the assays detect as few as 100 copies of template per RT reaction with excellent assay efficiency and linearity. We will also dis- cuss the appropriate uses of positive controls to normalize results between assays and between experiments, and how the EPIK RNA Spike-In controls can be used to build an accurate picture of the relative concentration of miRNA in a sample. These controls can be particularly effective in addressing complex biological problems such as the abundance of miRNA in exosomes from human blood.
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