Quencher extension for single nucleotide polymorphism quantification in bacterial typing and microbial community analyses.
Quencher extension is a novel single-step closed tube real-time method to quantify single nucleotide polymorphisms (SNPs) in combination with primer extension. A probe with a 5'-reporter is single-base extended with a dideoxy nucleotide containing a quencher if the target SNP allele is present. The reaction is measured from the quenching (reduced fluorescence) of the reporter. The relative amount of a specific SNP allele is determined from the nucleotide incorporation rate in a thermocycling reaction. The quencher extension protocol presented was developed for SNP allele quantification in Listeria monocytogenes and for microbial community analyses.
Rudi
Knut
K
Matforsk AS, Norwegian Food Research Institute, Oslo, Norway.
Zimonja
Monika
M
Skånseng
Beate
B
D015373:Bacterial Typing Techniques; D017931:DNA Primers; D017753:Ecosystem; D006801:Humans; D008089:Listeria monocytogenes / Q000145:classification* / Q000235:genetics / Q000254:growth & development / Q000472:pathogenicity; D016133:Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D017422:Sequence Analysis, DNA
D017931:DNA Primers