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However, we strongly recommend performing a qPCR assay to determine the optimal number of PCR cycles for the library amplification. The PCR Add-on and Reamplification Kit for Illumina V2 (Cat. No. 020.96208) includes the PCR and Mix “PM”, the PCR Enzyme Mix reagents and P7 primer (7000) “PE” and “P7” and “P5” primers that are required for the qPCR assay. In addition, SYBR Green I Nucleic Acid Gel Stain (10,000X in DMSO, Sigma-Aldrich Cat. No. S9430) is also required (user provided).
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What do I need for the qPCR Assay?
How do I calculate the Endpoint PCR cycle number from the qPCR assayendpoint cycle numbers?