Minimally-invasive liquid biopsies facilitate expedient diagnosis and monitoring of diseases, as well as opportunity for the early detection of diseases. Liquid biopsy assays utilize low volumes of serum, plasma and other biofluids in order characterize cell-free DNA (cfDNA). Circulating cfDNA are small DNA fragments (~150-200 bp) found in biofluids. Healthy human levels of plasma cfDNA are < 50 ng/mL, but often disease states are associated with higher levels of cfDNA. In fact, the release of cfDNA is attributed to apoptotic and necrotic tumor cells or active DNA release by tumor cells, and cancer-specific genetic variants can be identified from cfDNA. The methylation status of cfDNA can also be correlated to the presence of specific diseases and has shown utility as a diagnostic biomarker. Moreover, methylated cfDNA is a promising noninvasive prenatal testing marker given the differences between maternal peripheral blood and fetal or placental DNA

cfDNA Bisulfite Sequencing
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Using ORB's bisulfite sequencing service, methylation patterns can be examined from circulating cfDNA that is derived from blood-based biofluids. ORB scientists have developed protocols to investigate global and targeted methylation signatures. Both approaches are optimized to minimize biofluid or cfDNA input and to promote the conservation of precious samples.
Specific genomic regions, such as CpG islands, can be monitored using bisulfite sequencing through the selection of specific DNA sequences in the prepared libraries using high density capture reagents from Roche, Illumina, and Agilent.
Unlock the epigenetic profile of your biofluid samples with ORB’s bisulfite sequencing service!

cfDNA Bisulfate Sequencing Bioinformatic Deliverables
gDNA Read Summary
  • Detected sequence reads with genomic coordinates, counts per sample, and annotations.
Standard cfDNA Bisulfate Sequencing Report
  • Identification of methylated sites showing the methylation call and percentage methylation at each site.
  • Statistical analysis results showing methylated loci with significant differences in methylation levels between groups, annotation of nearest gene, and the relative position of the methylated site to the gene.
  • Maps of CpG-rich genomic regions indicating the distribution of deferentially methylated regions across the chromosomes.
  • Distribution histograms of CpG coverage and CpG methylation levels.
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ORB offers cost-effective examination of oncogenes from tumor tissue, biofluids, or genomic or cfDNA samples using targeted sequencing of oncopanels developed by industry leading manufacturers such as Illumina and Swift Biosciences. Only panels with expert–defined content are utilized in order to identify variants and fusion events with established relevance to cancer genetics. Extraction services are optimized for multiple samples types including biofluids to facilitate examination of circulating genetic variants collected with minimally invasive strategies.

Standard cfDNA Bisulfate Sequencing Report
  • Input requirements as low as 10 ng of genomic or cfDNA
  • Compatible with degraded DNA and FFPE tissue sections
  • Hematological malignancies , solid tumor, and pan-cancer panels available
  • Comprehensive coverage of all coding regions of the TP53 gene available with many panels
  • Examine genetic variants at low allelic frequencies (< 1%)
    • Germline and somatic mutations
    • Insertions/deletions (INDELS)
    • Single nucleotide polymorphisms
    • Loss of heterozygosity
    • Gene amplification
Data Analysis Deliverables
  • Known and potentially novel variants
  • Confidence scores and frequencies of detected variants
  • Functional annotation of each identified variants Data Analysis Deliverables