We’re redefining next-generation sequencing (NGS) capabilities and opening advanced applications so you can further your research. Our proprietary DNBSEQ technology provides extremely efficient and accurate real PCR-free NGS.

How does DNBSEQ compare to
traditional methods?

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NGS Test no-index-hopping-1940x1100

Higher Accuracy – Lower cost – More efficient

FeatureDNBSEQPCR clones
Clonal errorsNoneYes
Indel errors3x less
Ability for non-UMI applicationsYesNo
Index hoppingNoneYes
Ultra-dense patterned arrayAllowedNo
Ultra-higher throughput arrayYesNo
Occupany on flow cell~95%60-70%
qPCR requiredNoYes
Duplication rate<2%Much higher
See DNBSEQ in Action
DNBSEQ Graphic
NGS Testings DNBSEQ-technology-workflow-1940x835
NGS Testing
How Does our Sequencing Technology Work?

Called DNBSEQ, our core technology is built around DNA nanoballs (DNBs). Created during library prep, compact DNBs are loaded onto flow cells and the sequence is read with four fluorescent probes recognizing four DNA bases. DNBSEQ then uses lasers to excite the probes, while taking millions of images to identify bases. These sharp and bright images are then fed into our proprietary image analysis algorithms to precisely sequence each sample.

Complete Genomics - Kits and Reagent
Library Preparation

During sequencing library prep, the single-stranded circular DNA (sscirDNA) molecules are created. Typical double-stranded DNA fragments, with adapter sequences at the terminal ends, are heated to generate ssDNA. A splint oligonucleotide is hybridized to both ssDNA ends to form nicked circles, and a DNA ligase repairs the nick to create complete single-stranded circles.

With the single-stranded circle as a template, DNBSEQ uses rolling circle replication (RCR) to create billions of DNA nanoballs (DNBs) in a single tube. Each copy is made from the original DNA circle, eliminating clonal amplification errors, and reducing the GC biases and dropouts, often produced by PCR. Under proprietary conditions, the concatmer with about 300-500 template copies is folded into a tiny DNA nanoball ~200nm in diameter. 

Complete Genomics - Sequencing Platforms

The DNBs are loaded onto flow cells, which are etched with a pattern of  uniformly-spaced ~200nm binding sites at submicron distances. Each site bind single DNB, ensuring high yield of accurate reads with sharp and bright signal and no interference from neighboring nanoballs. 

The flow cell loading requires no expensive DNA quantification instruments or reagents. There is no under- or over-loading in a wide range of DNB concentrations. Proprietary loading buffers ensure DNBs stick to the same spots for hundreds of cycles maintaining strong signals. These refinements enable exceptional sequencing accuracy with minimal reagent consumption.

From there, combinatorial probe-anchor synthesis (cPAS) chemistry hybridizes sequencing primers to the DNBs and fluorescently labeled reversibly terminated probes are incorporated by a proprietary DNA polymerase in consecutive sequencing cycles. The fluorescent probes are then excited by laser light and the DNB array is imaged using advanced cameras.

After completing the first strand, the second, complementary strand attached to the original DNB is synthesized by controlled MDA and sequenced. This generates a stronger second strand signal with high sequencing accuracy.

Complete Genomics - Software and Analysis

Base calls are determined, and a quality score is calculated from four signal intensities. Our sub-pixel array registration algorithms extract precise fluorescent intensities per each DNB. GPU-accelerated and optimized algorithms, and real-time image analysis dramatically increase data processing speed and base-calling accuracy.

Explore our full suite of solutions

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