Whole Exome Sequencing with Next-Generation Sequencing

With a workflow requiring less than 50 minutes of hands-on time, the Ion AmpliSeq Exome RDY Kit enables enrichment and library construction in less than 6 hours. The kit requires less hands-on time compared to other exome enrichment technologies (Figure 1), and as little as 50 ng of DNA can be used.

The efficiency of exome enrichment is measured by specificity, defined as the percent of on-target bases, and coverage uniformity, defined as the percent of bases covered at a certain sequencing depth. High uniformity in sequence coverage is important for exome sequencing since this decreases the amount of sequencing needed to achieve a desired coverage threshold (e.g., 20x). Coverage depth is important for accurate variant calling. The Ion AmpliSeq Exome RDY Kit enables high efficiency of enrichment with >90% on-target bases and >90% coverage uniformity (Figure 2).

Other bench-top sequencing platforms are not as flexible, and running a small number of samples may be costly or researchers may have to wait for large batches of samples to fill each run. With the scalable chip format of the Ion GeneStudio S5 Series, you can cost-efficiently sequence one to four exomes per run without the need to batch samples. Simply select between the Ion 540 and Ion 550 chips (whichever fits your project size). For higher throughput laboratories, sequence up to 48 exomes in a working week* using Ion Chef System, Ion GeneStudio Prime System, and Ion 550 Chip.

The Ion AmpliSeq exome research solution integrates software and a bioinformatics pipeline to deliver a focused list of annotated exome variants, which helps to reduce the need for expensive bioinformatics resources. 

Torrent Suite Software is preconfigured to support Ion AmpliSeq exome sequencing including optimized variant calling parameters, enabling you to discover relevant variants with confidence. Torrent Suite Software provides a simple run quality report, access to coverage metrics, and full variant calling—all without the need for any command line skills.

Ion Reporter Software enables data to be easily uploaded via a small, efficient file directly from Torrent Suite Software into Ion Reporter Software workflows—allowing researchers access to public annotations of called variants and simple, intuitive filters of only the most relevant variants.

For more complicated experiments, like pairs or trios, more sophisticated Ion Reporter workflows are available to find variants unique to one sample in either a pair or a trio analysis.