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xGen® Duplex Seq Adapters—Tech Access

High accuracy detection of ultra-low frequency variants

The xGen Duplex Seq Adapters—Tech Access provide the highest level of error correction for next generation sequencing (NGS) applications. Dual-strand molecular barcoding is used to distinguish true variants from sequence errors that are introduced during sample preparation or sequencing. Improved error correction increases sensitivity and specificity, leading to greater accuracy and more confident reporting of rare and subclonal mutations detection, cell-free DNA (cfDNA) sequencing, drug resistance monitoring, and other applications.

xGen Duplex Seq Adapters—Tech Access are designed and optimized for use on Illumina platforms.

  • Achieve accurate detection of variants below 0.1% allele frequency using UMI-aware error correction
  • Analyze challenging, clinically relevant samples, such as FFPE and cfDNA
  • Easily incorporate adapters into common library prep and target enrichment workflows

Ordering

Sample indexing primers

Indexed primers for incorporating P5 and P7 adapter sequences.

Custom designs

For information on custom adapter designs, please contact Application Support at applicationsupport@idtdna.com.

The xGen Duplex Seq Adapters—Tech Access contains an equimolar pool of duplexed adapters, with each strand containing a 3 bp unique molecular identifier (UMI; Figure 1). The adapters are compatible with T-A ligation–based library preparation kits. The adapter pool is normalized to 2 nmol, delivered in a tube, and provided at a concentration of 15 µM in 30 mM HEPES, pH 7.5; 100 mM potassium acetate. Nuclease-Free Duplex Buffer (catalog # 11-01-03-01) is recommended for dilution, if necessary.

xGen Duplex Seq Adapters—Tech Access does not include the full Illumina P5 and P7 adapter sequences or sample indexes necessary to generate sequencing-ready libraries. P5 and P7 sample indexing primers must be ordered separately using the sample indexing primer order form. These primers, each bearing a unique index, must be used in PCR amplification with xGen Duplex Seq Adapters to establish full-length adapters that include dual UMIs and sample indexes.

Use xGen Duplex Seq Adapters—Tech Access with xGen Universal Blockers–TS Mix to improve on-target capture during targeted sequencing.

Figure 1. xGen Duplex Seq Adapters—Tech Access enable use of genetic information embedded in both DNA strands. xGen Duplex Seq Adapters—Tech Access are designed to be compatible with standard library construction methods. The adapters contain 3′-dTMP overhangs that are ligated to 3′-dA-tailed inserts. xGen Duplex Seq Adapters—Tech Access incorporate degenerate unique molecular identifiers (UMIs) to tag double-stranded molecules in the ligation step and unique, dual sample indexes are incorporated by PCR amplification. During post-sequencing analysis, the top and bottom strands of the original DNA input molecules can be paired back together to help identify errors introduced in the workflow.

NGS Tech Access Program

IDT is committed to providing our customers with high performance products for researchers working on the cutting edge of scientific discovery. The NGS Tech Access program at IDT is intended to accelerate innovation by enabling earlier access to our most advanced genomics tools. However, Tech Access products are not functionally QC-tested, so no certificates of analysis are available. These products are particularly well suited to experienced researchers who require the most up-to-date technology to unlock new discoveries.