An alternate nonisotopic technique of PCR-mediated allele-specific oligonucleotide analysis for the detection of a point mutation.
We report a simple protocol for the allele analysis of a point mutation using polymerase chain reaction. Conditions have been determined to use probes prepared for the conventional allele-specific oligonucleotide hybridization method directly. After amplification of the genomic region containing the mutation site, the nested polymerase chain reaction was performed using an allele-specific oligonucleotide probe as one side of the primer. Specificity of the amplification was achieved by decreasing the primer concentration in a two segment thermal cycling pattern. This protocol does not require additional special primers and an allelic mutation can be determined non-isotopically.
Nakajima
H
H
Second Department of Internal Medicine, Osaka University Medical School, Suita, Japan.
Hamaguchi
T
T
Tomita
K
K
Uenaka
R
R
Yamagata
K
K
Hanafusa
T
T
Kuwajima
M
M
Noguchi
T
T
Tanaka
T
T
Kono
N
N
D000483:Alleles; D001483:Base Sequence; D004247:DNA / Q000235:genetics; D015342:DNA Probes; D005784:Gene Amplification; D006801:Humans; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D009841:Oligonucleotides / Q000235:genetics*; D017354:Point Mutation; D016133:Polymerase Chain Reaction
D015342:DNA Probes; D009841:Oligonucleotides; D004247:DNA