In this Application Note, we focus on HiFi sequencing of full-length PCR amplicons spanning the 3.3 kb CYP21A2 and its pseudogene, CYP21A1P. PacBio HiFi reads are long (up to 25 kb) and accurate (99.9%) making them well suited to analyze the complexity of CAH-related pathogenic variants, where gene conversions and chimeric genes are the predominant mutation types.
PacBio® HiFi reads are long (up to 25 kb) and accurate (99.9%). Targeted HiFi sequencing of amplicons has the ability to span entire genes, allowing for straightforward haplotype construction, detection of structural variants or copy number variants in addition to SNVs and indels. These attributes make targeted HiFi sequencing well suited to analyze both rare and common variants of all types, thus increasing the positive detection rate of thalassemia-related variants.
With PacBio® HiFi sequencing, CYP2D6 variation can be captured using highly accurate long reads on a single platform. Amplicon-based HiFi sequencing fully and directly resolves and phases complex loci like CYP2D6 without assembly or inference, allowing for ancestry agnostic haplotype assignment.
The HiFiViral SARS-CoV-2 kit is an end-to-end solution
for genomic surveillance of COVID-19 variants. The kit
utilizes molecular inversion probes (MIPs), a
differentiated enrichment technology which combines
the simplicity and cost-effectiveness of PCR with the
resilience to mutations of target capture. The MIPs
approach, coupled with PacBio HiFi reads and SMRT®
Link software analysis, delivers several advantages
over PCR amplicon sequencing