DNA nucleic acid sequence-based amplification-based genotyping for polymorphism analysis.
Nucleic acid sequence-based amplification (NASBA) is a sensitive isothermal transcription-based amplification method known to be a suitable tool for RNA research. We demonstrate that NASBA technology can be applied to single nucleotide polymorphism (SNP) analysis using human genomic DNA as a template. Combination of DNA NASBA with multiplex hybridization of specific molecular beacons makes it possible to unambiguously discriminate the presence of the SNP of interest. This protocol is easy-to-use, robust, and makes it possible to rapidly detect single nucleotide substitutions in clinical or cell line DNA sequences using a large range of DNA input. Such a real-time genotyping DNA NASBA assay can find broad application in clinical diagnostics.
Berard
Cecile
C
bioMérieux, Marcy l'Etoile, France.
Cazalis
Marie-Angélique
MA
Leissner
Philippe
P
Mougin
Bruno
B
D001483:Base Sequence; D004247:DNA / Q000032:analysis* / Q000302:isolation & purification; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques / Q000295:instrumentation / Q000379:methods*; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D012313:RNA / Q000032:analysis* / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods
D012313:RNA; D004247:DNA