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.
CITATION STYLE
Guo, D. C., & Milewicz, D. M. (2007). Universal primer applications for pyrosequencing. Methods in Molecular Biology (Clifton, N.J.), 373, 57–62.
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