Pyrosequencing: History, biochemistry and future
2006 (English)In: Clinica Chimica Acta, ISSN 0009-8981, E-ISSN 1873-3492, Vol. 363, no 02-jan, 83-94 p.Article, review/survey (Refereed) Published
Background: Pyrosequencing is a DNA sequencing technology based on the sequencing-by-synthesis principle. Methods: The technique is built on a 4-enzyme real-time monitoring of DNA synthesis by bioluminescence using a cascade that upon nucleotide incorporation ends in a detectable light signal (bioluminescence). The detection system is based on the pyrophosphate released when a nucleotide is introduced in the DNA-strand. Thereby, the signal can be quantitatively connected to the number of bases added. Currently, the technique is limited to analysis of short DNA sequences exemplified by single-nucleotide polymorphism analysis and genotyping. Mutation detection and single-nucleotide polymorphisin genotyping require screening of large samples of materials and therefore the importance of high-throughput DNA analysis techniques is significant. In order to expand the field for pyrosequencing, the read length needs to be improved. Conclusions: Th pyrosequencing system is based on an enzymatic system. There are different current and future applications of this technique.
Place, publisher, year, edition, pages
2006. Vol. 363, no 02-jan, 83-94 p.
pyrosequencing, real-time monitoring, SNP, bioluminscence, dna-binding-protein, polymerase-chain-reaction, single-nucleotide polymorphisms, micromachined filter-chamber, allele-specific extension, real-time pyrophosphate, tumor-suppressor gene, i reaction pathway, firefly luciferase, escherichia-coli
IdentifiersURN: urn:nbn:se:kth:diva-15369DOI: 10.1016/j.cccn.2005.04.038ISI: 000234535600007ScopusID: 2-s2.0-28644434277OAI: oai:DiVA.org:kth-15369DiVA: diva2:333410
QC 201005252010-08-052010-08-05Bibliographically approved