Ion AmpliSeq™ HD Library Kit with HD Enhancer

The Ion AmpliSeq HD Library Kit with HD Enhancer is designed for the manual generation of Unique Molecular Tag (UMT)-targeted amplicon libraries for use as part of the Ion AmpliSeq HD sequencing workflow. This product includes both an Ion AmpliSeq HD Library Kit (Cat. No. A37694) and an HD Enhancer kit for improved library quality and molecular coverage results. It is compatible with custom panels created using the Ion AmpliSeq Designer tool (www.ampliseq.com) and sequenced using an Ion GeneStudio S5 Series instrument.

Ion AmpliSeq HD technology enables scalable polymerase chain reactions (PCRs) from 12- to 5000-plex (1 x 5000-plex in a 1-pool panel or 2 x 2500-plex in a 2-pool panel) for UMT-targeted sequencing, with a level of detection down to 0.1% variant frequency depending on the sample type, sample input amount, and read coverage depth.

Key product features include:
• Custom design creation
• Ultra-high sensitivity detection
• Dual-barcoded sample multiplexing
• Bi-directional sequencing
• Improved library preparation performance

Custom design creation
The Ion AmpliSeq Designer tool can be used to create custom designs for use with Ion AmpliSeq HD technology. The designs are made-to-order and allow the user the flexibility of creating region, hotspot, or fusion designs. These are one- or two-pool designs recommended for use with samples obtained from cfDNA, cfRNA, cfTNA, FFPE DNA, FFPE RNA, gDNA, or total RNA.

Ultra-high sensitivity detection
Ion AmpliSeq HD technology is capable of ultra-high levels of detection down to 0.1%. These detection levels have been observed for cfDNA, cfTNA, and gDNA controls with known variants using an input amount of ≥ 20 ng at adequate read coverage depth. Levels of detection for other sample types, such as FFPE, will vary.

Dual-barcoded sample multiplexing
The Ion AmpliSeq HD Library Kit with HD Enhancer includes reagents for generating amplicons using Ion AmpliSeq HD primers, which are used to prepare libraries for sequencing. The libraries are dual-barcoded at the 5’ and 3’ ends using the Ion AmpliSeq HD Dual Barcode Kit 1-24, which enables the multiplexing of up to 24 samples in a single chip.

Bi-directional sequencing
The Ion AmpliSeq HD Library Kit also provides sequencing information from both strands. This enables better error correction and higher confidence in the quality of your sequence data.

Improved library preparation performance
The new HD Enhancer component included in this kit, in association with an updated Ion AmpliSeq HD library preparation protocol, improves the product performance by reducing primer-dimer formation during library preparation and improving amplicon performance, especially high-GC amplicons molecular coverage.