Pyrosequencing: history, biochemistry and future.
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 polymorphism 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.
Ahmadian
Afshin
A
Department of Biotechnology, Royal Institute of Technology (KTH), Stockholm, Sweden.
Ehn
Maria
M
Hober
Sophia
S
D002624:Chemistry, Clinical / Q000379:methods* / Q000639:trends; D004247:DNA / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D003933:Diagnosis; D005838:Genotype; D006801:Humans; D008156:Luciferases / Q000378:metabolism; D008164:Luminescent Proteins / Q000737:chemistry; D009942:Organometallic Compounds / Q000737:chemistry; D009943:Organophosphorus Compounds / Q000737:chemistry; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D017422:Sequence Analysis, DNA / Q000379:methods* / Q000639:trends; D013430:Sulfate Adenylyltransferase / Q000378:metabolism; D013698:Templates, Genetic
D008164:Luminescent Proteins; D009942:Organometallic Compounds; D009943:Organophosphorus Compounds; C066850:Cascade Blue; D004247:DNA; D008156:Luciferases; D013430:Sulfate Adenylyltransferase; D004259:DNA-Directed DNA Polymerase