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Pyrosequencing is a method of DNA sequencing (determining the order of nucleotides in DNA) based on the "sequencing by synthesis" principle, in which the sequencing is performed by detecting the nucleotide incorporated by a DNA polymerase. Pyrosequencing relies on light detection based on a chain reaction when pyrophosphate is released. Hence ...
English: Gray, blue, red, green, light green, black graph papers with 1 cm–0.5 cm–1 mm grids (page size: A4) in printable PDF format. Date 25 July 2013, 18:04:17
Standard flowgram format (SFF) is a binary file format used to encode results of pyrosequencing from the 454 Life Sciences platform for high-throughput sequencing.SFF files can be viewed, edited and converted with DNA Baser SFF Workbench (graphic tool), or converted to FASTQ format with sff2fastq or seq_crumbs.
A challenge of short-read sequence data is particularly an issue in sequencing new genomes (de novo) and in sequencing highly rearranged genome segments, typically those seen of cancer genomes or in regions of chromosomes that exhibit structural variation.
Pyrosequencing uses luciferase to generate light for detection of the individual nucleotides added to the nascent DNA, and the combined data are used to generate sequence reads. [81] This technology provides intermediate read length and price per base compared to Sanger sequencing on one end and Solexa and SOLiD on the other. [90]
In a de Bruijn graph, there is a possibility of 4^k different nodes to make arrangements of a genome. The number of nodes used to create the graph can be reduced in number by considering only the k-mers found within the DNA strand of interest. Given sequence 1, it is possible to determine the nodes of size 7, or 7-mers, that will be in the graph.
Pyrosequencing, a new method of DNA sequencing which involves the addition of phosphorylated dNTP's to the DNA polymerase reaction which emmit light upon binding to the DNA template. It makes DNA sequencing significantly faster than with the current chain termination method.
Screenshot of a chromatogram inside the program "Sequencher" Capillary Electrophoresis to Electropherogram process (Courtesy of www.biointeractive.org) Generation of results