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Vega targeted datasets

Targeted HiFi sequencing datasets with the Vega benchtop system

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Explore targeted panels on the Vega benchtop system

Uncover the full picture of your region of interest with highly accurate HiFi long-read targeted sequencing, now in-house with Vega benchtop system. The unique combination of long, highly accurate HiFi reads and deep target coverage delivers a powerful approach for resolving complex loci, revealing the full spectrum of variation, including structural variants and, with PureTarget technology, methylation changes in the same assay.

From carrier screening to characterizing gene editing outcomes and complex cancer genome rearrangements, targeted HiFi sequencing is transforming translational research and clinical testing. If repetitive regions, complex targets, unclear phasing, difficult amplification, or missing methylation data are limiting your assay performance, targeted HiFi sequencing provides the clarity, completeness, and confidence needed for reliable clinical insights.

 

Highlights:

 

Targeted HiFi methods

Struggling with repetive regions?

BERRY GENOMICS LEADS THE WAY WITH AMPLICON SEQUENCING FOR GENETIC DISEASE RESEARCH

One of China’s more prominent clinical genomics companies, Berry Genomics, is a leader in the prevention of genetic birth defects. Berry achieved the first regulatory clearance of a clinical-grade long-read sequencer when the Sequel II CNDx system received Class III Medical Device Registration approval from the National Medical Products Administration (NMPA) in China. Berry was also the first customer to receive shipment of the benchtop Vega systems, which join Berry’s fleet of HiFi platforms. Berry has sequenced more than 300,000 Thalassemia samples and helped contribute to a 3-5% increase in findings compared to traditional assays.

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“With HiFi long-read sequencing, we found at least 20 novel large deletions or structural variations in the Thalassemia genes that have helped contribute to a 3–5% increase in diagnosis rate compared to traditional assays.”

– Dr. Aiping Mao, Vice Director of R&D at Berry Genomics

Need clear and accurate haplotype phasing?

Targeted hybrid capture combined with HiFi long-read sequencing panels enables clear and accurate haplotype phasing across complex genomic regions. The long range and high accuracy of HiFi reads allow variants to be resolved on the same molecule, delivering confident phase determination without ambiguity from repetitive or GC-rich sequences. The result is a more complete and clinically meaningful view of inherited and disease-associated variation.

 

Hybrid capture app brief     Agilent CGP panel and PacBio HiFi poster

Trouble amplifying your target region?

Learn how gene editing researchers are using PureTarget to characterize engineered animal models with exceptional accuracy. With high depth of coverage of native DNA, it is possible to identify mosaicism in founder lines and track inheritance in their progeny.

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Capture methylation and variants in the same assay?

PureTarget uses the CRISPR-Cas9 system to generate targeted native DNA libraries. This amplification-free approach retains epigenetic signals like methylation and avoids PCR artifacts.

With streamlined workflows, scalable multiplexing, and deep coverage of difficult regions, PureTarget panels replace multiple assays and give labs the power to profile challenging loci like FMR1, FXN, F8, and SMN1 in a single assay.

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Carrier panel

A curated panel of hard-to-sequence genes, built to ACMG guidance and designed to reduce the need for add-on assays.

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Repeat expansion panel 2.0

A comprehensive panel of 38 neurological disease targets, including 17 ataxia genes and targets for ALS/FTD, Huntington disease, and myotonic dystrophy.

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Custom control panel

A set of three control targets for designing and benchmarking custom panels. Includes X-linked UBL4A and autosomal ACTB and GAPDH.

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Comprehensive genotyping of the most challenging targets

Repeat expansions in the FXN and RFC1 genes occur in high frequency across many human populations, with recent estimates that 1 in ~14 individuals carry premutations at RFC1 and 1 in ~100 carry pre-mutations at FXN (Ibañez et al., 2024). Accurate repeat sizing is important to identify carriers of pathogenic alleles longer than 70 repeats for FXN and longer than 400 repeats at RFC1 (Leitão et al., 2024). Long and accurate HiFi reads make it possible to distinguish among the diverse repeat motifs at RFC1, only some of which are pathogenic (Dominik et al., 2023). 

RFC1 expansions

TRVZ plot of NA20752 at RFC1(ital) repeat showing consensus allele for the normal (top) and pathogenic expanded allele (bottom) with phased reads aligned to each consensus sequence. 657 “AAGGG” motifs are observed in the expanded allele with consensus length of 3253 bp.
TRGT plot: NA20752 at RFC1 repeat showing consensus allele for the normal (top) and pathogenic expanded allele (bottom) with phased reads aligned to each consensus sequence. Also, 657 “AAGGG” motifs are observed in the expanded allele with consensus length of 3253 bp.

 

TRVZ plot of HG01175 at RFC1 repeat showing consensus allele for the normal (top) and pathogenic expanded allele (bottom) with phased reads aligned to each consensus sequence. 393 “AAGGG” motifs are observed in the expanded allele with consensus length of 1948 bp. Note the diversity of repeat motifs which are distinguishable by HiFi reads.
TRGT plot: HG01175 at RFC1 repeat showing consensus allele for the normal (top) and pathogenic expanded allele (bottom) with phased reads aligned to each consensus sequence. Also, 393 “AAGGG” motifs are observed in the expanded allele with consensus length of 1948 bp. Note the diversity of repeat motifs which are distinguishable by HiFi reads.

 

FXN expansions

Long allele Short allele
Sample Coverage Observed motif count Expected motif count Coverage Observed motif count Expected motif count
HM16212 92 471 500 146 8 <30
NA16212 68 515 500 99 8 <30
NA16202 49 817 830 86 8 <30
NA16237 49 699 700 134 8 <30

High on-target coverage enables accurate genotyping

The PureTarget repeat expansion panel enables deep coverage for accurate genotyping of repeat expansions associated with human disease. The table below highlights typical coverage for Coriell samples with known repeat expansions. Note that 2 µg of DNA was prepared with the PureTarget repeat expansion panel of 20 targets, sequenced on the Vega system, and analyzed with TRGT (Dolzhenko et al., 2024) in the SMRT Link PureTarget repeat expansion panel analysis workflow. Full dataset available here.

 

Sample Target with repeat expansion Total reads per sample Mean target coverage Coverage of expanded allele Epanded allele size (bp)
20 gene panel
HM23709 AR 54,252 343 182 147
NA13716 ATN1 40,700 234 88 216
NA13536 ATXN1 39,845 267 132 124
NA06153 ATXN3 64,164 258 114 207
ND14442 C9orf72 16,821 51 4 4563
HM03756 DMPK 52,504 395 282 1305
NA09237 FMR1 36,167 257 113 2683
NA16215 FXN 67,939 245 62 2963
NA13509 HTT 58,870 480 225 252
HM23629 PABPN1 50,569 417 218 27
HG01175 RFC1 53,454 378 57 1948

Sample to insight workflow

Extraction

Start with HMW DNA extracted from human blood or cell lines using Nanobind extraction kits.

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Library prep

Targeting and library prep can be completed in 8 hours. All reagents supplied in the kit.

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Sequencing

Sequence up to 48 samples per Vega SMRT Cell with 24 hour movies and optimized run conditions for PureTarget libraries.

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Data analysis

SMRT Analysis is automatically configured to generate library QC, target enrichment statistics, and tandem repeat genotypes using TRGT (Dolzhenko et al., 2024).

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What can you expect from one sequencing run on the Vega system?

Targeted panels Samples per Vega SMRT Cell
PureTarget panels 96
Amplicon sequencing > 1,000
Hybrid capture
20 Mb panel 12
2 Mb panel 72
100 kb panel 288
Manual library prep with PureTarget 24 kit (103-707-900) and assumes >100x per target.

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References

Dolzhenko, E., et al. (2024). Characterization and visualization of tandem repeats at genome scale. Nat Biotechnol. 2024 doi: 10.1038/s41587-023-02057-3.

Dominik, N., et al. (2023). Normal and pathogenic variation of RFC1 repeat expansions: implications for clinical diagnosis. Brain. 146(12):5060-5069. doi: 10.1093/brain/awad240.

Ibañez, K., et al. (2024) Increased frequency of repeat expansion mutations across different populations. Nature Medicine. https://doi.org/10.1038/s41591-024-03190-5

Leitão, E., et al. (2024). Identification and characterization of repeat expansions in neurological disorders: Methodologies, tools, and strategies. Rev Neurol (Paris). 180(5):383-392. doi: 10.1016/j.neurol.2024.03.005.

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