Genotyping pyrosequencing

Van Booven, D. J. (2006) Pyrosequencing genotype storage techniques. Methods Mol Biol. 373, 177-186. 

Dideoxynucleotides are commercially available with different fluorescent tags therefore, the resulting extension products can be examined using laser scanners to see which fluorophore has been incorporated. This method directly reveals the genotype of the template DNA. The method can be multiplexed and is commercially available (SNaPshot, see below). Alternatively to the use of dideoxynucleotides, regular deoxynucleotides can be used sequentially in the extension reaction. Here, the primer is essentially extended as in a regular PCR reaction. The only difference is the sequential addition of one nucleotide at a time. This method is commercially available (see pyrosequencing, described below). 

Saeki, M., Saito, Y., Jinno, H., et al. (2003) Comprehensive UGTIAI genotyping in a Japanese popnlation by pyrosequencing. Clin. Chem. 49, 1182-1185. 

Garsa, A., S. Marsh, S., and McLeod, H. L. CYP3A4 and CYP3A5 genotyping by pyrosequencing. BMC Medical Genetics. In press. 

Nordfors, L., Jansson, M., Sandberg, G., et al. (2000) Large-scale genotyping of single nucleotide polymorphisms by Pyrosequencing and validation against the 5 nuclease (Taqman assay. Hum. Mutat. 19, 395 01. 

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