Analyzing the millions of disease research samples stored in biorepositories around the world is a primary unmet need for biomedical researchers. Sequencing these samples using next-generation sequencing (NGS) has been difficult, as typically only nanogram amounts of DNA can be isolated, far less than current commercial protocols require. In addition, NGS creates a flood of data, requiring expensive bioinformatics expertise to interpret. The latest set of Life Technologies Ion AmpliSeq™ and Ion Reporter™ products, when augmented with specific and sensitive confirmation using TaqMan® assays, offer turnkey solutions to overcome these challenges.
The introduction of the Ion PGM™ Sequencer democratized sequencing—it is a fast, simple, scalable, and affordable platform. The Ion AmpliSeq Cancer Panel, Comprehensive Cancer Panel, and Inherited Disease Panel further expand the capabilities of this platform by transforming oncology and disease research.
Ion AmpliSeq panels, whether predesigned and ready to use, or custom-built to probe specific genes of interest, deliver a simplified single-day workflow comprising target selection, amplification, sequencing, and analysis (Figure 1). Further, with the use of Ion Reporter Software, the integrated workflow also includes an annotated readout detailing the biological significance of observed gene mutations. The Ion AmpliSeq technology requires only tens of nanograms of input DNA. Orthogonal confirmation of variants observed with Ion AmpliSeq panels is accomplished by selecting from more than 4.5 million ready-to-use TaqMan assays or by designing a custom TaqMan assay.
TaqMan assays deliver high sensitivity and specificity for SNP genotyping, and they may be analyzed in standard or digital PCR mode if increased sensitivity is required to detect low-frequency or somatic mutations.
Targeted sequencing with Ion AmpliSeq technology is scalable in multiple ways: multiplexing of primer pairs in each tube, multiple Ion Chips with different throughputs, and up to 96 barcodes for sample multiplexing. The number of primer pairs per pool is scalable, allowing researchers to target anything from a single gene to hundreds of genes while maintaining a low number of pools per panel.