pyrosequencing


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pyrosequencing

a real-time DNA sequencing technique based on monitoring DNA synthesis by BIOLUMINESCENCE using four enzymes: DNA POLYMERASE, ATP sulfurylase, LUCIFERASE and apyrase. The NUCLEOTIDES are tested one at a time for incorporation into the nascent DNA strand during synthesis. Each incorporation event, by DNA polymerase, generates pyrophosphate (PPi) as a by-product. The PPi is converted, by ATP sulfurylase, to ATP and this drives the conversion of luciferin to oxyluciferin by luciferase, with the emission of a light pulse. The light produced is detected by a charge coupled device camera and displayed as a peak in a light profile (pyrogram(tm)). Peak height relates to the number of nucleotides incorporated. If not incorporated the nucleotide is degraded by apyrase and no light is emitted. The DNA sequence can be determined from the pyrogram, knowing the sequence of nucleotides added.
Collins Dictionary of Biology, 3rd ed. © W. G. Hale, V. A. Saunders, J. P. Margham 2005
References in periodicals archive ?
Goel, "Pyrosequencing enhancement for better detection limit and sequencing homopolymers," Biochemical and Biophysical Research Communications, vol.
A total of 302,327 valid sequences of 16 samples were obtained after 454 pyrosequencing and 158,015 high quality sequences after filtering and removing chimera, in which the most abundant sequences were obtained from the sample DQ, and the least were from the sample CY, as shown in Table 1.
Table 1--Detection of LAM resistance mutations by direct sequencing and pyrosequencing. Source LAM resistance M1 2429 1181 mutations detection HBV-RNA Direct YMDD YMDD YVDD sequencing Pyrosequencing YMDD (94%) YMDD YMDD (30%) YIDD (6%) YVDD (70%) HBV-DNA Direct YMDD YMDD YVDD sequencing Pyrosequencing YMDD (96%) YMDD YVDD YIDD (4%)
The methylation patterns of CDKN2A, FOXA1, HOXD8, OCT4, SOX2, and SOX17 were evaluated by pyrosequencing. As shown in Figure 1, no significant differences were observed when all tested patient groups were evaluated.
Pyrosequencing. Initial pyrosequencing PCR assays with the validation primers described above for B.
This has implications for barcoding pyrosequencing studies which cover high sample numbers, because less barcoded PCR amplicons per sample will be detected and, therefore, dominant bacteria will play a larger role in determining the bacterial composition.
A total of 299 ant samples were subjected to 454 pyrosequencing and combined with data from 11 samples analyzed by Ishak et al.
The aim of this study was to describe the fungal communities present in fecal samples obtained from healthy dogs and dogs with acute, nonhemorrhagic diarrhea using high-throughput 18S rRNA gene pyrosequencing.
DNA was then subjected to 454 pyrosequencing, a "fingerprinting" technique that provides a snapshot of all the bacterial types present.
We also discuss the limitations of the Sanger DNA sequencing technique in quality control testing of pDNA and suggest using the more sensitive DNA pyrosequencing technique.