DIRECT DETECTION OF SINGLE-NUCLEOTIDE POLYMORPHISMS IN BACTERIAL DNA BY SNPtrap

作者:Gronlund Hugo; Moen Birgitte; Hoorfar Jeffrey; Radstrom Peter; Malorny Burkhard; Rudi Knut*
来源:Preparative Biochemistry & Biotechnology, 2011, 41(2): 166-174.
DOI:10.1080/10826068.2011.547366

摘要

A major challenge with single-nucleotide polymorphism (SNP) fingerprinting of bacteria and higher organisms is the combination of genome-wide screenings with the potential of multiplexing and accurate SNP detection. Single-nucleotide extension by the minisequencing principle represents a technology that both is highly accurate and enables multiplexing. A current bottleneck for direct genome analyses by minisequencing, however, is the sensitivity, since minisequencing relies on linear signal amplification. Here, we present SNPtrap, which is a novel approach that combines the specificity and possibility of multiplexing by minisequencing with the sensitivity obtained by logarithmic signal amplification by polymerase chain reaction (PCR). We show a SNPtrap proof of principle in a model system for two polymorphic SNP sites in the Salmonella tetrathionate reductase gene (ttrC).

  • 出版日期2011

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