The optimal number of PCR cycles for a given input amount of total RNA can vary depending on sample quality and origin. The optimal cycle number for a specific sample type should be determined using the qPCR assay.
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the PCR Add-on and Reamplification Kit V2 for Illumina (Cat. No. 208)
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.
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The recommended protocol for the qPCR assay is provided in the PCR Add-on and Reamplification Kit V2 User Guide.
Additional Resources
For additional information on the qPCR assay, consult the following online FAQs: https://faqs.lexogen.com/faq/pcr-add-on-and-reamplification-kit-v2