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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© 2007 Humana Press Inc.
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Guo, DC., Milewicz, D.M. (2007). Universal Primer Applications for Pyrosequencing®. In: Walker, J.M., Marsh, S. (eds) Pyrosequencing® Protocols. Methods in Molecular Biology™, vol 373. Humana Press. https://doi.org/10.1385/1-59745-377-3:57
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DOI: https://doi.org/10.1385/1-59745-377-3:57
Publisher Name: Humana Press
Print ISBN: 978-1-58829-645-0
Online ISBN: 978-1-59745-377-6
eBook Packages: Springer Protocols