Do I need to perform a qPCR assay to determine the number of PCR cycles for the EP PCR?

We strongly recommend performing a qPCR assay to determine the optimal number of PCR cycles for the endpoint PCR.

To perform the qPCR assay you will need:

• miRVEL Profiling PCR Add-on and Reamplification Kit (Cat. No. 246), and

• SYBR Green I nucleic acid stain (10,000X in DMSO, not provided in the kit. We recommend using Sigma-Aldrich, Cat. No. S9430 or ThermoFisher, Cat. No. S7585)

ATTENTION! The use of SYBR Green I-containing qPCR mastermixes from other vendors is not recommended.

NOTE: The recommended protocol for the qPCR assay is provided in the PCR Add-on and Reamplification Kit User Guide.