Universal primer applications for pyrosequencing.
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.
Guo
Dong-Chuan
DC
Department of Internal Medicine, University of Texas Medical School ot Houston, Houston, TX, USA.
Milewicz
Dianna M
DM
D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics* / Q000378:metabolism; D011756:Diphosphates / Q000378:metabolism*; D006801:Humans; D016133:Polymerase Chain Reaction; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D017422:Sequence Analysis, DNA / Q000379:methods*
D017931:DNA Primers; D011756:Diphosphates