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PubMed · 17185757

Universal primer applications for pyrosequencing.

Dong-Chuan Guo·Dianna M Milewicz
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Abstract

Pyrosequencing is a high-throughput technique for single-nucleotide polymorphism (SNP) genotyping, DNA sequencing, and SNP allele frequency and DNA methylation assays. The Pyrosequencing assay involves initially purifying PCR-generated single-strand DNA labeled with biotin as a sequencing template. The disadvantage of the synthesis of a biotinylated sequence-specific primer to assay each DNA variant lies in costs and time consumption. To overcome this problem, methodology has been developed to generate biotinylated Pyrosequencing DNA templates using a universal biotinylated primer. Using this methodology, biotinylated DNA fragments can be generated without the use of sequence-specific biotinylated primers for each DNA variant.

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Base SequenceDNA PrimersDiphosphatesHumansPolymerase Chain ReactionPolymorphism, Single NucleotideSequence Analysis, DNAJournal ArticleResearch Support, N.I.H., ExtramuralResearch Support, Non-U.S. Gov't

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Dong-Chuan Guo, Dianna M Milewicz. 2007. Universal primer applications for pyrosequencing.. https://doi.org/10.1385/1-59745-377-3%3A57

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