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Next Generation Sequencing Services

Advanced Genomic Insights with Next-Generation Sequencing

We offer high-throughput sequencing using second and third generation sequencing and optical genome mapping platforms to deliver high-quality, interpretable results. 


RNA Sequencing (RNA-Seq)

 

Our RNA-Seq platforms are compatible with a variety of sample types, including cell lines, snap-frozen tissue, whole peripheral blood, FFPE blocks and sections, peripheral blood mononuclear cells (PBMCs), total RNA, and biopsy tissue. Compatible library preparation kits include non-Illumina for mRNA, Illumina Truseq for mRNA, and ribosomal RNA-seq for mRNA, lncRNA, and microRNA. 

Applications: 

  • Differential gene expression 
  • Functional annotation, pathway and network analysis 
  • Driver mutation prediction 
  • Tumor purity analysis 
  • Molecular subtyping analysis 
  • Comparative analysis with published human tumor samples upon request 
  • Options to accommodate other analyse

rna-sequencing

 

Whole Exome Sequencing (WES)

 

Our WES service is suitable for sequencing-based analysis of cell lines, snap-frozen tissue, whole peripheral blood, FFPE blocks and sections, PBMCs, biopsy tissue, and mouth swabs. 

Applications: 

  • Identify genomic variants on DNA level, such as gene mutation, and copy number variants 
  • Co-expression network analysis 
  • Comparative analysis with published human tumor samples upon request 
  • Options to accommodate other analyses 

whole-exome-sequencing

 

Whole Genome Sequencing (WGS) / Whole Genome Bisulfite Sequencing (WGBS)

 

WGS/WGBS platforms are compatible with multiple sample types, including cell lines, snap-frozen tissue, whole peripheral bloodFFPE blocks and sections, PBMCs, biopsy tissue, and mouth swabs. 

WGS applications: 

  • Gene mutation identification 
  • Copy number variants 
  • Large structural variants 

wgs

WGBS applications: 

  • Epigenomics 
  • Gene methylation analysis

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Single Cell Sequencing (10x Genomics)

 

Our single cell sequencing service is suitable for analysis of cell lines, fresh tissue, whole peripheral blood, and PBMCs in single cell RNA-Seq and T cell receptor (TCR) sequencing assays 

Applications: 

  • Characterize and identify heterogeneous cell populations 
  • Discover new cell markers and regulatory pathways 
  • Uncover novel cell types, cell states & rare cell types 
  • Reconstruct developmental hierarchies and reveal lineage relationships 
  • Profiling healthy and diseased tissue and organs 

We also offer bioinformatics analysis for our single cell sequencing service, which includes: 

  • QC and cell/gene filtering 
  • Cluster analysis on integrated samples 
  • Exploration of known cell type markers 
  • Identification of cluster-specific markers 
  • Functional analysis on cluster-specific markers 
  • Further customized analysis upon request 

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Third Generation Sequencing – PacBio Sequel II Platform

 

Crown Bioscience uses the PacBio Sequel II platform, which offers single-molecule, real-time (SMRT) sequencing. Long reads are readily assembled into complete genomes for sequencing of full-length transcripts with >99.999% consensus accuracy. Other benefits include uniform coverage of typically inaccessible regions, single-molecule resolution, and direct detection of epigenetic modifications. 

Applications: 

  • WGS—de novo assembly, variant detection, and structural variant detection 
  • RNA sequencing—full-length transcripts for whole transcriptome and genome annotation 
  • Complex populations—full-length 16S, metagenomic functional profiling, and shotgun metagenomic assembly 

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Optical Genome Mapping (OGM) - Bionano Saphyr® Platform

 

Unrivaled Structural Variant Detection with Bionano Genomics Saphyr® Platform

Large structural variations are responsible for many diseases and conditions, including cancers and developmental disorders. Optical genome mapping with Saphyr detects structural variations ranging from 500 bp to megabase pairs in length and offers assembly and discovery algorithms that far outperform sequencing-based technologies in sensitivity.

  • The Bionano Saphyr® System detects structural variations in an unbiased manner at much higher sensitivities than sequencing-based technologies, and routinely at 5% variant allele fraction

  • Complementing OGM with sequencing can reveal more answers to challenging genomic questions in genetic diseases and cancer research

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Key Applications:

  • For cancer research:
    • Genome variation common to cancer is too complex for low coverage whole genome sequencing
    • Complex rearrangements as well as highly repetitive regions of the genome present additional challenges for sequencing
    • For heterogeneous cancer samples, Saphyr detects all types of structural variants down to 1% variant allele fraction
  • For genetic disease:
    • Find new candidate genes
    • Identify new variants in known genes
    • Discover repeat expansions
    • Finds variants other technologies can’t see
  • Genome assembly:
    • Reveal true long-range structure of the genome while reducing sequencing costs
    • Build-in pipeline “hybrid scaffold” could increase the accuracy of genome assembly
    • Including OGM data into de novo genome assemblies has become a gold standard
  • Advance cytogenomics:
    • Replace traditional cytogenetics with convenience, higher resolution and a more complete picture of complex aberration.
    • 10,000x greater resolution compared to karyotyping
    • Consolidate traditional cytogenetic assays into a single workflow
Method Resolution Features
Karyotyping -5-10 Mbp Extensive training required for interpretation, slow, labor-intensive data collection and analysis.
Cell culture required.
FISH -100 kbp Targeted, extremely limited approach; only shows handful of variants; slow, labor-intensive data
collection. Requires validation of every lot of probes utilized
Array based
techniques
-50 kbp Cannot detect balanced rearrangements. Cannot resolve nature of a structural aberration.
OGM with
Bionano Saphyr
500 bp Fully automated detection of: CNVs, repeat expansion, FSHD, unbalanced events from
single exon level to aneuploidies, balanced events, inversions, translocations,
gene fusions down to 1% VAF.

Microbiome Analysis  

 

The PacBio Sequel II and Illumina Novaseq 6000 platforms for microbiome culture and analysis are compatible with feces, DNA, tissue and microbiome samples.  

Applications: 

  • Full-length 16S rRNA sequencing with PacBio Sequel II platform) 
  • Metagenomic sequencing
  • Metatranscriptomic sequencing

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Nanostring Services

 

We utilize the nCounter® platform for Nanostring analysis.  

  • Array-based multiplex analysis of up to 800 RNA, DNA, or protein targets
  • Flexible sample types including FFPE, tissue, lysates, and biofluid samples
  • Panels available:
  • PanCancer IO 360 Panel
  • PanCancer Immune Profiling Panel
  • PanCancer Pathways Panel
  • PanCancer Progression Panel
  • Check other available panels here

Applications : 

  • Gene expression analysis 
  • Solid tumor profiling 
  • Immuno-oncology profiling 
  • Gene fusion analysis 
  • Single cell gene expression analysis 
  • miRNA expression analysis 
  • Copy number variation analysis 
  • lncRNA expression analysis 
  • ChIP-String expression analysis 

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