The rhAmpSeq system enables highly accurate amplicon sequencing on Illumina® next generation sequencing (NGS) platforms. Whether you are investigating thousands of targets or a few, the fast and easy rhAmpSeq workflow, based on our proprietary RNase H2-dependent PCR (rhAmp PCR) technology, generates NGS-ready amplicon libraries for deep, targeted resequencing.
The rhAmpSeq system offers a simple, easy workflow with only 2 PCR amplification steps, using either a custom or predesigned panel. To begin creating your amplicon libraries, simply choose a panel, then add the rhAmpSeq Library Kit and rhAmpSeq Index Primers.
The rhAmpSeq system offers consistent performance across panel sizes ranging from tens to thousands of amplicons in a single multiplex reaction (Figure 3), and supports a variety of applications, including challenging sample types [e.g., formalin-fixed paraffin-embedded (FFPE) and cell-free DNA (cfDNA) samples]. Table 2 summarizes key features, specifications, and performance metrics for the rhAmpSeq system.
The data shown in Figure 3 are representative of performance from 3 different custom rhAmpSeq panels using random SNP markers in the human genome with initial (non-optimized) rhAmpSeq panel designs. The performance of your custom panel may depend on several factors, including the quality of the input samples and the reference genome in the case of non-human species.
Table 2. rhAmpSeq system features and specifications.
|Supported protocols||Regular library preparation (10–100 ng)|
High-throughput library preparation (10–50 ng)
|Sample type||Tissue, FFPE, cfDNA|
|Insert size||Flexible (50–200 nt)|
|Custom panel size||Up to 5000 amplicons per panel|
|Sample indexing capability||96 index sequences (up to 9216 combinations)|
Figure 3. High quality sequencing data across a range of non-optimized panel sizes and DNA input quantities. Coriell DNA samples were used to evaluate the performance of non-optimized rhAmpSeq panels of varying sizes (20, 282, and 994 amplicons) following the high-throughput (HT) protocol using 10 and 50 ng of DNA input. The largest panel (994 amplicons) was evaluated following the regular (Reg) protocol with 10 and 100 ng of DNA input. Coverage uniformity is the percent of targets with coverage ≥0.2X of the mean. Error bars = standard deviation from the mean.
The rhAmpSeq system can be used in a wide variety of research applications, and should be considered for any application that requires stringent and efficient sequencing analysis. These are just a few of the fields in which rhAmpSeq technology has been successfully used.
Detecting disease-associated variants is a key element of human-focused research. With the rhAmpSeq system’s combined specificity and multiplexing capabilities, you can easily and accurately detect “hotspot” variants. In addition, the rhAmpSeq Sample ID Panel can be used to track samples and avoid mix-ups in human sample workflows.
Create highly multiplexed amplicon sequencing panels for your targets of interest.
Predesigned amplicon sequencing panel for tracking and managing human samples in NGS analysis workflows.
Rapid, cost-effective library preparation for rhAmpSeq targeted sequencing.
Easily prepare and combine rhAmpSeq libraries onto a single sequencing run for maximum efficiency.