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*Input range for QuantSeq V2 3’ mRNA-Seq FWD is 1 - 500 ng of total RNA. Cycle numbers for QuantSeq Flex depend entirely on the targeting strategy and should always be optimized using the qPCR Assay.
ATTENTION! The cycle number ranges below above are not recommendations for the given input amounts for Endpoint PCR! These ranges are provided as a guideline only for choosing an approximate number of cycles for the Endpoint PCR Test. The ranges account for the 3 extra cycles needed when using 1.7 µl of cDNA for the Endpoint PCR Test, versus 17 µl (10x more) that is used for standard Endpoint PCR.
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The PCR Add-on and Reamplification Kit V2 (Cat. No. 208), 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.
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