The PCR Add-on kit contains a PCR Mix including an Illumina P5 specific primer, a thermostable polymerase, and an Illumina P7 primer without a barcode referred to as Barcode 00 for 96 PCR reactions. By adding SYBR Green I to the PCR reaction a qPCR assay to determine the optimal number of cycles for the endpoint PCR of your QuantSeq or SENSE cDNA libraries can be determined.

The qPCR assay is recommended to determine the exact number of cycles for the endpoint PCR in order to prevent any under- or overcycling of your library. Undercycling may result in too little library while overcycling can lead to significant distortions in gene expression values.

The kit furthermore contains a reamplification primer that can be used to reamplify already barcoded libraries if they were undercycled to get enough material for sequencing.

NOTE: By 17.02.2017 the protocol was adjusted to be compatible with the QuantSeq upgrade, which includes the renaming of BC00 to P7 Primer 7000.
•Kit size: 96 rxn
•Compatibility information ◦Product: SENSE mRNA-Seq V2, SENSE Total RNA-Seq, and QuantSeq
◦Platform: Illumina