The PCR Add-on kit contains a PCR Mix including a non-indexed Illumina P5 specific primer, a thermostable polymerase, and an Illumina P7 primer without a barcode (primer 7000). In addition, a reamplification primer for single-indexed (i7 only ) is included. 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 CORALL cDNA libraries can be determined.The kit can be used for two purposes:
Reamplification of single-indexed QuantSeq and CORALL libraries
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
Reamplification of already barcoded libraries can be performed if they were undercycled, in order to get enough material for sequencing.
NOTE: This kit is not compatible with the small RNA-Seq Library Prep Kit.