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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 ...
These 7-mers then create the graph shown in figure 1. [1] The graph shown in figure 1 is a very simple version of what a graph could look like. [2] This graph is formed by taking the last 6 elements of the 7-mer and linking it to the node whose first 6 elements are the same.
The pyrosequencing method is based on the detection of the pyrophosphate release on nucleotide incorporation. Before performing pyrosequencing, the DNA strand to sequence has to be amplified by PCR. Then the order in which the nucleotides have to be added in the sequencer is chosen (i.e. G-A-T-C).
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]
Similar to bisulfite sequencing and pyrosequencing, this method quantifies methylation levels at various loci within the genome. This assay is used for methylation probes on the Illumina Infinium HumanMethylation27 BeadChip (henceforth, 27k [methylation] array).
Dye-primer sequencing facilitates reading in an optical system for faster and more economical analysis and automation. The later development by Leroy Hood and coworkers [7] [8] of fluorescently labeled ddNTPs and primers set the stage for automated, high-throughput DNA sequencing. Sequence ladder by radioactive sequencing compared to ...
Pyrosequencing has also been used to analyze bisulfite-treated DNA without using methylation-specific PCR. [7] [8] Following PCR amplification of the region of interest, pyrosequencing is used to determine the bisulfite-converted sequence of specific CpG sites in the region. The ratio of C-to-T at individual sites can be determined ...
A ternary plot, ternary graph, triangle plot, simplex plot, or Gibbs triangle is a barycentric plot on three variables which sum to a constant. [1] It graphically depicts the ratios of the three variables as positions in an equilateral triangle .
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