Lexogen, the transcriptomics and next-generation sequencing company, announced the launch of the QuantSeq-Pool Sample-Barcoded 3′ mRNA-Seq Library Prep Kit. The QuantSeq-Pool method combines sample barcoding and early pooling with the benefits of Lexogen’s established QuantSeq 3′ mRNA-Seq kit, which has been already cited in more than 550 publications. Hence, QuantSeq-Pool tremendously increases sample multiplexing capacities in gene expression profiling projects, saving time and costs.
To date, gene expression profiling is the most commonly used application for RNA-Seq in biomedical research and diagnostics. Conventional gene expression projects use mRNA sequencing technologies that offer comprehensive single-nucleotide resolution across the full-length transcript but are limited in the number of samples that can be processed and multiplexed cost-efficiently and economically. Although the sequencing price per base is continuously decreasing, sample preparation, sequencing, and data processing are still major cost factors that limit high-throughput screenings.
3′ mRNA-Seq methods, like Lexogen’s widely-used QuantSeq protocol, generate Next-Generation Sequencing libraries close to the 3′ end of polyadenylated RNAs without the need of prior poly(A) enrichment. Since QuantSeq 3′ mRNA-Seq generates only one fragment per transcript, the number of reads mapped to a given gene is directly proportional to its expression level. Thus, QuantSeq facilitates data analysis by using simple counting and thereby offers a reliable and affordable solution for gene expression profiling.
Lexogen’s new QuantSeq-Pool kit accelerates gene expression profiling studies by enabling sequencing of up to 36,864 RNA samples in parallel. During the initial library generation step, sample barcodes are introduced allowing subsequent pooling of the individually labeled samples. Such batch processing significantly shortens the complete RNA-to-Sequencing workflow by reducing handling time and eliminating the need for quantification, individual quality control, and equimolar pooling of individual libraries. During the library amplification step additional Unique Dual Indices (UDIs) can be introduced for higher multiplexing capacity. Thus, triple-indexed QuantSeq-Pool libraries (containing i1 sample-barcodes and Lexogen’s 12 nt UDIs) are highly suitable for large-scale screening projects.
“QuantSeq set the gold standard for 3′ sequencing. Even though its speed and simple workflow was unmatched to begin with, the early pooling approach of QuantSeq-Pool will on the one hand greatly reduce time and overall costs, but also further increase data quality by reducing technical variability. The development of this kit was strongly driven by customer demand and QuantSeq-Pool is setting a new standard for high throughput gene expression profiling” said Alexander Seitz, Lexogen’s founder and CEO.
Source – Lexogen