The QuantSeq 3’ mRNA-Seq Library Prep method generates Next Generation Sequencing (NGS) libraries from single sites close to the poly(A) tail of RNAs. Due to its fast and highly sensitive workflow this tag profiling method is ideal for the accurate quantification of gene expression, also from low-input and degraded samples. QuantSeq has been widely used for diverse studies and features in more than 150 peer-reviewed publications.
“Here we quantified mRNA expression from 299 maize lines that represent the genotypic and phenotypic diversity of modern inbred maize. We automated a 3′ mRNA sequencing method (QuantSeq, Lexogen GmbH), which is more efficient and accurate than mRNA sequencing and deals well with paralogues.” Kremling, K. A. G. et al. (2018) Dysregulation of expression correlates with rare-allele burden and fitness loss in maize. Nature. DOI: 10.1038/nature25966.
The determination of differential gene expression is a standard application and often complemented by gene set enrichment (GSEA) and pathway analyses to derive the network of differentially activated genes. Other applications include the determination of alternative polyadenylation sites and their differential usage, the 3’ (re)annotation of genes, and polysome profiling. The QuantSeq-Flex version is applied to profile non-polyadenylated RNA and to build custom gene expression profiling panels.
QuantSeq libraries are intended for a high degree of multiplexing. With the barcodes provided up to 9,216 libraries can be sequenced on an Illumina lane. This high level of multiplexing allows saving costs as the length restriction in QuantSeq saves sequencing space.
The straightforward procedure and the tremendously reduced sequencing depth requirements make the method highly cost efficient, and by adding robustness, sensitivity and versatility it is no wonder that QuantSeq has established itself as the most widely applied 3’ RNA-Seq protocol.