Long, Processive Enzymatic DNA Synthesis Using 100% Dye-Labeled Terminal Phosphate-Linked Nucleotides

Korlach, Jonas; Bibillo, Arek; Wegener, Jeffrey; Peluso, Paul; Pham, Thang; Park, Insil; Clark, Sonya; Otto, Geoff; Turner, Stephen W.

doi:10.1080/15257770802260741

Cited by 78 publications

(44 citation statements)

References 21 publications

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“…By forgoing the need to use a locus-specific approach, next generation sequencing, in principle, permits the identification of mutations arising anywhere in the genome. In the near future, third generation DNA sequencing technologies will be available that allow high processivity reads of single strands of DNA [Korlach et al, 2008]. This will permit the analysis of highly repetitive regions of DNA that are likely to be very variable, opening up a new dimension to studies of mutation.…”

Section: The Promise Of Genomics Technologiesmentioning

confidence: 99%

Germ cell mutagens: Risk assessment challenges in the 21st century

Singer

Yauk

2010

Environ and Mol Mutagen

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Heritable mutations may result in a wide variety of detrimental outcomes, from embryonic lethality to genetic disease in the offspring. Despite this, today's commonly used test batteries do not include assays for germ cell mutation. Current challenges include a lack of practical assays and concrete evidence for human germline mutagens, and large data gaps that often impede risk assessment. Moreover, most regulatory assessments are based on the assumption that somatic cell mutation assays also protect the germline by default, which has not been adequately confirmed. The field is also faced with new challenges aimed at dramatically reducing animal testing, and attempts to rapidly classify thousands of chemicals using high throughput in vitro assays. These approaches may not adequately capture effects that may be particular to gametes, since many aspects of the germline are unique. In light of these challenges, an urgent need exists to develop new approaches to evaluate the potential of toxicants to cause germline mutation. The application of new technologies will greatly enhance our understanding of mutation in humans exposed to environmental mutagens. However, we must be poised to collect and interpret these data, and facilitate risk translation to regulators and the public. Genetic toxicologists must also become actively involved in the development of high-throughput tools to study germline mutation. Appropriate attention to these areas will result in the development of policies that prioritize the protection of the germline and future generations from DNA sequence mutations.

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Section: The Promise Of Genomics Technologiesmentioning

confidence: 99%

Germ cell mutagens: Risk assessment challenges in the 21st century

Singer

Yauk

2010

Environ and Mol Mutagen

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“…Recently, a novel promising sequencing method, Single-Molecule, Real-Time (SMRT) DNA sequencing technology, was announced (http://www.pacificbiosciences.com/index.php?q=home). It is based on eavesdropping on a single DNA polymerase molecule working in a continuous, processive manner [80] . SMRT DNA sequencing can read long sequences in short time with high quality data and low cost, but it is still in development and may not be available in market for several years.…”

Section: Impact Of New Sequencing Technologies On Bacterial Genome Rementioning

confidence: 99%

Genomic research for important pathogenic bacteria in China

et al. 2009

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Rapid accumulation of bacterial genomic data offered an unprecedented opportunity to understand bacterial biology from a holistic view of point. We can thus closely look at the way in which a pathogen is evolved, and these data has been applied to molecular epidemiology and microbial forensics, and screening of novel diagnostic, vaccine and drug targets. The newly developed high-throughput low-cost sequencing technologies, such as 454, Solexa and SOLiD, will promote the acquisition and application of genomic data in new research areas that we dared not imagine previously, such as the metagenomics of human gastric-intestinal tract, for better and comprehensive understanding of human health and disease.

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“…A single Ф-29 DNA polymerase enzyme molecule, a highly processive and strand displacing enzyme, is immobilized in a small hole called zero-mode wave guide (ZMW) to process the extension of a single molecule of primed DNA template (Eid et al, 2009). Four color phospholinked dye labeled nucleotides are used in this process and their fluorescence is quenched until they are incorporated during the sequencing reaction (Korlach et al, 2008). In the ZMW, as each nucleotide is incorporated by the anchored DNA polymerase, the phospholinked dye label is cleaved and its fluorescence light pulses are captured by four single photon sensitive cameras in the sequencing instrument (Lundquist et al, 2008).…”

Section: Pacific Biosciences Rs -Smrt™ (Single Molecule Real Time) Sementioning

confidence: 99%