The QuantSeq Kit is a library preparation protocol designed to generate Illumina and Ion Torrent compatible libraries of sequences close to the 3’ end of the polyadenylated RNA.
QuantSeq kits are available in two versions. QuantSeq FWD generates NGS reads towards the poly(A) tail, directly reflecting the mRNA sequence. It is offered for both Illumina and Ion Torrent platforms. QuantSeq Reverse (REV) is only available for Illumina platforms and requires a Custom Sequencing Primer (CSP Version 2, included in the kit) for Read 1. With QuantSeq REV and the CSP it is possible to exactly pinpoint the 3’ end in Read 1 as the first nucleotide of your NGS read corresponds the very last nucleotide of the mRNA. The reads generated during Read 1 reflect the cDNA sequence
The required input amount of total RNA is as low as 100 pg (QuantSeq for Illumina) and 5 ng (QuantSeq for Ion Torrent). QuantSeq is suitable to reproducibly generate libraries from low quality RNA, including FFPE samples
QuantSeq libraries are intended for a high degree of multiplexing. External barcodes allowing up to 96 samples to be sequenced per lane on an Illumina flow cell are included in QuantSeq 3’ mRNA-Seq Library Prep Kits for Illumina. In-line barcodes allowing up to 48 samples to be sequenced in one sequencing run of Ion Torrent instruments are included in QuantSeq 3’ mRNA-Seq Library Prep Kit for Ion Torrent. This high level of multiplexing allows saving costs as the length restriction in QuantSeq saves sequencing space. QuantSeq is also designed to yield insert sizes for short sequencing reads (SR50, SR100).