MEET THE NEW KITS
What is PureTarget?
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.
Amplification-free target enrichment workflow
- Start with high molecular weight DNA extracted from human blood or lymphoblastoid cells with Nanobind kits
- Dephosphorylate to block DNA ends
- Cut DNA with Cas9 and pair of guide RNAs on each side of target
- Add dA tail
- Ligate indexed SMRTbell adapters
- Remove non-SMRTbell templates with nuclease digestion
- Pool and sequence 96 samples on Revio + SPRQ with automation kit (8-48 samples on Vega or Revio + SPRQ with manual kit)
Puretarget Datasets
Purchasing guide for PureTarget Reagents
Modular bundles give flexibility with content and scale
Playlist
SEGMENT SPOTLIGHT: CLINICAL RESEARCH PURETARGET- PCR-FREE ENRICHMENT WITH METHYLATION FOR CHALLENGING GENES
SEGMENT SPOTLIGHT: CLINICAL RESEARCH PURETARGET- PCR-FREE ENRICHMENT WITH METHYLATION FOR CHALLENGING GENES
Consolidate your lab’s efforts by replacing multiple, labor-intensive assays with a single, scalable PureTarget panel that screens 20 targets and up to 48 samples at once. Our PCR-free workflow provides a complete characterization of tandem repeats by accurately sizing long expansions, sequencing interruption motifs, and detecting methylation status in one assay. By sequencing native DNA without PCR, the PureTarget workflow eliminates GC dropout and amplification bias, ensuring you get the most accurate and reliable data for even the most challenging genomic regions.
Carrier panel variant calling
The PacBio PureTarget Carrier Pipeline is a WDL-based workflow designed to genotype tandem repeat regions and homologous genes with segmental duplications using PacBio PureTarget HiFi data. It is available in the DNA Nexus marketplace, Golden Helix VarSeq, and deployable to on-prem servers from GitHub.
Paraphase: Genotyping highly similar paralogous genes
For hundreds of medically important genes like SMN1 and CYP21A2, highly similar paralogs and pseudogenes create genomic blind spots, making variant calling challenging. In this installment of the Spotlight on Human Genomics series, PacBio bioinformatics scientist Xiao Chen introduces Parapahse, a toll designed to resolve these complex regions.
Profiling variation in and around tandem repeats with TRGT
Tandem repeats have long been a blind spot for researchers, despite their link to many neurological diseases and contributing a huge amount of variation to our genomes. In this installment of the Spotlight on Human Genomics series, PacBio bioinformatics scientist Egor Dolzhenko introduces TRGT, the Tandem Repeat Genotyping Tool – a new approach designed to comprehensively resolve variation in tandem repeat regions.
Explore
PacBio has over 10,000 articles, reports, papers, and videos related to genomic sequencing
PureTarget FAQs
PureTarget is amplification-free so the best use cases for PureTarget include:
- Hard to amplify regions like tandem repeats or GC rich sequence
- Targets where methylation or phasing matters
- Genes with high sequence homology or large structural variants
Other advantage of PureTarget include:
- Automation scripts available for Hamilton NGS STAR MOA for PureTarget kit 96, 16 hour turn around time
- 8 hour turn around time with manual PureTarget kit 24
- Ability to combine multiple types of challenging targets into one panel (tandem repeat expansions, inversions, large deletions)
- Variant calling software pipelines available for repeat expansion panel and carrier panel
No problem! You can make in-silico (virtual) panels by masking out targets you don’t want. Simply modify your BED file to remove unwanted targets before analyzing in SMRT Link with Target Enrichment or PureTarget repeat expansion analysis. Only data from loci included in the BED file will be returned in the output files.
The officially supported sample types are Nanobind-extracted human blood, lymphoblastoid cells, and saliva (support for manual library prep and Revio SPRQ only), or Coriell lymphoblastoid cell DNA with GQN at 30 kb > 5. The best performance for PureTarget is with these samples, talk to your local rep or [email protected] about bundling Nanobind with your PureTarget kits.
- Download – Carrier panel coordinates (.XLS)
- Download – Repeat expansion panel 2.0 coordinates (.XLS)
- Download – Control panel coordinates (.XLS)
Also available from the application kit consumables page under each panel kit and in the hosted datasets
| CARRIER PANEL | |
|---|---|
| Gene | Disease |
| F8 | Hemophilia A |
| FXN | Friedreich ataxia |
| FMR1 | Fragile-X disease (FXS) |
| CYP21A2 | Congenital adrenal hyperplasia |
| TNXB | Classical-like Ehlers-Danlos syndrome |
| HBA1/2 | Alpha thalassemia |
| GBA | Gaucher disease |
| SMN1/2 | Spinal muscular atrophy |
| ARX | Early-infantile epileptic encephalopathy (EIEE1) and Partington syndrome (PRTS) |
| HBB | Sickle cell anemia and Beta thalassemia |
| RPGR | X-linked retinitis pigmentosa |
| AFF2 | Fragile X, FRAXE type |
| REPEAT EXPANSION PANEL 2.0 | |
|---|---|
| Genes | Disease |
| ATN1, ATXN1, ATXN2, ATXN3, ATXN7, ATXN8, ATXN10, CACNA1A, PPP2R2B, TBP | Spinocerebellar ataxia (SCA) |
| FMR1 | Fragile-X disease (FXS) |
| AFF2 | Fragile X syndrome, FRAXE type |
| AFF3 | Intellectual disability associated with fragile site FRA2A |
| C9orf72 | Frontotemporal dementia (FTD), amyotrophic lateral sclerosis (ALS) |
| FXN | Friedreich ataxia (FRDA) |
| RFC1 | Cerebellar ataxia, neuropathy, and vestibular areflexia syndrome (CANVAS) |
| NOTCH2NLC | Neuronal intranuclear inclusion disease, Alzheimer disease and parkinsonism phenotype (NIID) |
| DMPK, CNBP | Myotonic dystrophy (DM) |
| HTT | Huntington disease (HD) |
| JPH3 | Huntington’s disease-like type2 (HDL2) |
| TCF4 | Fuchs endothelial corneal dystrophy 3 (FECD3) |
| AR | Kennedy Disease, Spinal and bulbar muscular atrophy, (SBMA) |
| PABPN1 | Oculopharyngeal muscular dystrophy (OPMD) |
| ABCD3, GIPC1, LRP12, PILPL1 | Oculopharyngodistal myopathy (OPDM) |
| HOXD13 | Syndactyly (SD5) |
| PHOX2B | Congenital central hypoventilation syndrome (CCHS) |
| PRNP | Creutzfeldt-Jakob disease (CJD) |
| CSTB | Progressive Myoclonic Epilepsy Type 1 (EPM1) Unverricht-Lundborg Disease (ULD) |
| SAMD12 | Familial adult myoclonic epilepsy type 1 (FAME) |
| CONTROL PANEL | |
|---|---|
| Gene | Chromosome |
| UBL4A | chrX |
| ACTB | chr7 |
| GAPDH | chr12 |
