ArticleNucleic acids research2004
A multi-enzyme model for Pyrosequencing.
Article in Nucleic acids research, 2004. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed.
- 5-mC methylation study of sORFs in 3'UTR of transcription factor JUNGBRUNNEN 1-like during leaf rust pathogenesis in wheat.Molecular biology reports · 2024Article
- Structural insight into the assembly and working mechanism of helicase-primase D5 from Mpox virus.Nature structural & molecular biology · 2024Article
- The epigenetic modification of DNA methylation in neurological diseases.Frontiers in immunology · 2024Review
- Molecular testing for BRAF mutations to inform melanoma treatment decisions: a move toward precision medicine.Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc · 2018Review
- DNA and RNA analyses in detection of genetic predisposition to cancer.Hereditary cancer in clinical practice · 2012Article
- Transient model of thermal deactivation of enzymes.Biochimica et biophysica acta · 2011Article
- Mapping the cargo protein membrane translocation step into the PEX5 cycling pathway.The Journal of biological chemistry · 2009Article
- Structural optimization for heat detection of DNA thermosequencing platform using finite element analysis.Biomicrofluidics · 2008Article
- A multienzyme bioluminescent time-resolved pyrophosphate assay.Analytical biochemistry · 2007Article
- Analysis of read length limiting factors in Pyrosequencing chemistry.Analytical biochemistry · 2007Article
- Direct amplification of single-stranded DNA for pyrosequencing using linear-after-the-exponential (LATE)-PCR.Analytical biochemistry · 2006Article
- Estimating RNA editing efficiency of five editing sites in the serotonin 2C receptor by pyrosequencing.RNA (New York, N.Y.) · 2005Article
Corrections and comments
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Authors and funding
8 authors.
Funding
Abstract
Pyrosequencing is a DNA sequencing technique based on sequencing-by-synthesis enabling rapid real-time sequence determination. This technique employs four enzymatic reactions in a single tube to monitor DNA synthesis. Nucleotides are added iteratively to the reaction and in case of incorporation, pyrophosphate (PPi) is released. PPi triggers a series of reactions resulting in production of light, which is proportional to the amount of DNA and number of incorporated nucleotides. Generated light is detected and recorded by a detector system in the form of a peak signal, which reflects the activity of all four enzymes in the reaction. We have developed simulations to model the kinetics of the enzymes. These simulations provide a full model for the Pyrosequencing four-enzyme system, based on which the peak height and shape can be predicted depending on the concentrations of enzymes and substrates. Simulation results are shown to be compatible with experimental data. Based on these simulations, the rate-limiting steps in the chain can be determined, and K(M) and kcat of all four enzymes in Pyrosequencing can be calculated.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.