Rapid and sensitive detection of point mutations and DNA polymorphisms using the polymerase chain reaction.
We report a rapid and sensitive method for the detection of base changes in given sequences of genomic DNA. This technique is based on the facts that specific regions of genomic sequences can be efficently labeled and amplified simultaneously by using labeled substrates in the polymerase chain reaction and that in nondenaturing polyacrylamide gels, the electrophoretic mobility of single-stranded nucleic acid depends not only on its size but also on its sequence. The process does not involve restriction enzyme digestion, blotting, or hybridization to probes. We found that most single base changes in up to 200-base fragments could be detected as mobility shifts. RAS oncogene activation was detected by this technique. We also show that the interspersed repetitive sequences of human, Alu repeats are highly polymorphic.
Orita
M
M
Oncogene Division, National Cancer Center Research Institute, Tokyo, Japan.
Suzuki
Y
Y
Sekiya
T
T
Hayashi
K
K
D001483:Base Sequence; D002460:Cell Line; D004247:DNA / Q000235:genetics*; D004591:Electrophoresis, Polyacrylamide Gel; D005784:Gene Amplification; D011905:Genes, ras; D009154:Mutation; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D012091:Repetitive Sequences, Nucleic Acid
D004247:DNA