Quencher extension for single nucleotide polymorphism quantification in bacterial typing and microbial community analyses
Publikasjonsdetaljer
Tidsskrift : Methods in molecular biology , vol. 345 , p. 111–117–7 , 2006
Utgiver : Humana Press
Internasjonale standardnummer
:
Trykt
:
1064-3745
Elektronisk
:
1940-6029
Publikasjonstype : Vitenskapelig artikkel
Har du spørsmål om noe vedrørende publikasjonen, kan du kontakte Nofimas bibliotekleder.
Kjetil Aune
Bibliotekleder
kjetil.aune@nofima.no
Sammendrag
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.