Analysis of polymerase chain reaction product by capillary electrophoresis and its application to the detection of single base substitution in genes.
Capillary gel electrophoresis (CGE) was studied for the direct analysis of polymerase chain reaction (PCR) amplified samples. A low cross-linked polyacrylamide gel (3%T, 0.5%C) was used for CGE with treated and untreated silica capillaries. CGE showed high reproducibility and resolution in the separation of DNA fragments (ca. 100-1000 base pairs) produced by PCR. The CGE system was applied to the detection of an amplification refractory mutation system (ARMS) and PCR-restriction fragment length polymorphism (PCR-RFLP), which are detection methods of single base substitution in genes using PCR. With the CGE system, full automation of PCR product detection is feasible.
Arakawa
H
H
School of Pharmaceutical Sciences, Showa University, Tokyo, Japan.
Uetanaka
K
K
Maeda
M
M
Tsuji
A
A
D010584:Bacteriophage phi X 174 / Q000235:genetics; D004252:DNA Mutational Analysis / Q000379:methods*; D004279:DNA, Viral / Q000032:analysis*; D015252:Deoxyribonucleases, Type II Site-Specific; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D006801:Humans; D009154:Mutation; D010651:Phenylalanine Hydroxylase / Q000235:genetics; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length
D004279:DNA, Viral; D010651:Phenylalanine Hydroxylase; D015252:Deoxyribonucleases, Type II Site-Specific; C024403:GGCC-specific type II deoxyribonucleases