A mass screening device of genome by polymerase chain reaction-restriction fragment-single strand conformation polymorphism analysis.
We have developed a new genome screening method and named it as polymerase chain reaction-restriction fragment-single strand conformation polymorphism (PCR-RF-SSCP) analysis. This method consists of three steps: (1) amplification of long DNA by PCR; (2) digestion of the amplified PCR products by restriction enzyme(s) and labelling the restriction sites; (3) polyacrylamide gel electrophoresis under non-denaturating conditions (SSCP analysis) and under denaturating conditions containing 8M urea. Theoretically, this method enables us to detect even a base substitution, deletion or insertion in up to 22,000 base pairs amplified from genomic DNA by PCR in one analysis. The procedures are very simple, reproducible and require no special apparatus. This method is applicable to any genes already known and we believe that this method is very useful for mass screening of the genome.
Tawata
M
M
Third Department of Internal Medicine, University of Yamanashi Medical School, Japan.
Iwase
E
E
Aida
K
K
Onaya
T
T
D001483:Base Sequence; D017931:DNA Primers; D004272:DNA, Mitochondrial; D005820:Genetic Testing; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D018807:Polymorphism, Single-Stranded Conformational
D017931:DNA Primers; D004272:DNA, Mitochondrial