Targeted RNA sequencing library preparation: QuantSeq-Flex

The QuantSeq-Flex Kit from Lexogen enables the generation of Illumina®-compatible targeted RNA sequencing libraries from any RNA sample. Utilising custom primers for first or second strand synthesis, the QuantSeq-Flex kit features a simple workflow to generate ready-to-sequence libraries in just 4.5 hours. This kit includes additional modules for Reverse Transcription (RT) and Second Strand Synthesis (SSS) using custom primers and offers maximum flexibility of targets and input material.

Benefits of using the QuantSeq-Flex kit
• Direct counting for gene expression quantification: produces one fragment per transcript
• Sequence-ready libraries in just 4.5 hours, with <2 hours hands-on time
• Targeted RNA-Seq from any RNA sample
• Suitable for targeted sequencing of gene panels, enrichment detection of fusion genes, and various other types of transcripts
• Intended for a high degree of multiplexing
• Libraries are compatible with Illumina® sequencers
• No prior poly(A) enrichment or rRNA depletion required

QuantSeq-Flex targeted RNA-Seq library preparation workflow

Read 1 reflects the RNA sequence. The reverse transcription reaction can either be primed using an oligodT primer (included in the kit) or a target-specific primer (not included, custom designed for desired targets). Second strand synthesis can either be initiated by random priming (included in the kit) or by using a target-specific primer (not included, custom designed for desired targets). Depending on the combination of different priming options 4 different libraries can be generated.

Applications
• Gene expression studies of custom targets
• Targeted sequencing
• Molecular barcoding (UMI or inline indexing)

System compatibility
• HiSeq® 2000/2500/3000/4000
• NextSeq® 500/550/2000
• NovaSeq® 6000
• MiSeq®
• MiniSeq®
• iSeq®
• Genome Analyzer®

Material available for download
Application note: automation of QuantSeq 3' mRNA kit
QuantSeq-Flex first strand synthesis module V2 user guide
QuantSeq-Flex second strand synthesis module V2 user guide

QuantSeq-Flex citations

HiSeq, NextSeq, NovaSeq, MiSeq, MiniSeq, iSeq, Genome Analyzer and Illumina are a registered trademarks of Illumina