QuantSeq 3´mRNA-Seq Library Prep Kit for Ion Torrent
- QuantSeq kit:
- For gene expression and 3‘ UTR analysis
- 100 pg of total RNA input incl. from low-quality RNA and FFPE samples
- Cost-effective sequencing of up to 96 samples/lane
- 3´mRNA-Seq libraries in 4.5 hours
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.
Analysis of Low Quality Samples
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 maintains exceptional strand-specificity of >99.9% and allows to map reads to their corresponding strand on the genome, enabling the discovery and quantification of antisense transcripts and overlapping genes.
Simple Bioinformatics Analysis
Read mapping is simplified by skipping the junction detection. Reads are generated at the transcripts’ most 3′ end where nearly no junctions are located. Data processing can hence be accelerated by using e.g. Bowtie2 instead of TopHat2.
TheQuantSeq data analysis pipeline has been integrated on the Bluebee genomics analysis platform and is accessible for any user even without bioinformatics background. Learn more and get started at https://www.bluebee.com/quantseq/
QuantSeq’s simple workflow allows generating ready-to sequence-NGS libraries within only 4.5 hours, including less than 2 hours hands-on time.
Mapping of TES
QuantSeq allows to exactly pinpoint the 3’ end of poly(A) RNA and therefore obtain accurate information about the 3’UTR.
Perfect For Gene Counting
Just one fragment per transcript is produced; therefore no length normalization is required. This allows more accurate determination of gene expression values and renders QuantSeq the best alternative to microarrays and conventional RNA-Seq in gene expression and eQTL studies.
Cost Saving Multiplexing
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).