LUTHOR High-Definition Single-Cell 3′ mRNA-Seq Library Prep Kit
LUTHOR HD redefines the approach to single-cell RNA sequencing by offering an RNA-amplification-based kit able to detect virtually every single mRNA molecule present in the cell.
Like QuantSeq FWD, LUTHOR HD is a 3′ end mRNA-Seq kit that uses oligo(dT) primers to target polyadenylated RNAs and generate reads mapping to the 3′ end of transcripts. The two kits, however, use different protocols and technologies and are developed for different sample input amounts. While LUTHOR HD is designed for inputs ranging from 1 ng to 10 pg (equivalent to 100 cells to 1 cell) and even lower*, QuantSeq FWD is ideal for inputs ranging from 1 ng to 500 ng of total RNA.
LUTHOR HD is compatible with Lexogen’s patented 12 nt UDI Sets for maximal read rescue capabilities (no more misassigned or lost reads!).
The sensitivity of single-cell RNA-seq methods depends on multiple library preparation steps that may contribute to dropouts during reverse transcription, template-switching, tailing, or ligation. Any losses during these steps will result in reproducible data for highly expressed genes only. To compensate for low mRNA conversion rates, current high-throughput methods rely on sequencing large numbers of cells at shallow read depth, which provides an incomplete picture of gene expression. Although shallow sequencing is sufficient for cell type identification based on few highly expressed marker genes, it lacks the transcriptional complexity to define cell-specific heterogeneity.
LUTHOR HD overcomes sensitivity limitations by employing in vitro-mediated RNA amplification of poly(T)-tagged mRNA templates at the very beginning of the protocol, prior to conversion into NGS libraries.