Restriction fragment length polymorphism analysis of ERBB2 oncogene by capillary electrophoresis.
Capillary electrophoresis (CE) with a sieving buffer containing ethidium bromide was applied to the detection of PCR-amplified RFLP samples. With CE, in contrast to agarose gel electrophoresis, run times are short, i.e., typically less than 30 min, the capillary can be re-used, and full automation is feasible. The addition of ethidium bromide to the buffer system in conjunction with a field amplification injection technique led to increased sample detectability and resolution. Migration time precision was better than 0.2% RSD with a approximately 12-bp resolution for the DNA fragment sizes of interest. RFLP samples were analyzed for homo- or heterozygosity based on the presence of 500- and/or 520-bp DNA fragments. Special software was used to correct for run-to-run migration time variations, thus facilitating genotype assignment.
Ulfelder
K J
KJ
Beckman Instruments, Inc., Palo Alto, California 94304.
Schwartz
H E
HE
Hall
J M
JM
Sunzeri
F J
FJ
D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D015252:Deoxyribonucleases, Type II Site-Specific; D004587:Electrophoresis, Agar Gel; D006579:Heterozygote; D006720:Homozygote; D006801:Humans; D007438:Introns; D009857:Oncogenes; D016133:Polymerase Chain Reaction / Q000379:methods; D012150:Polymorphism, Restriction Fragment Length; D011505:Protein-Tyrosine Kinases / Q000235:genetics
D004247:DNA; D011505:Protein-Tyrosine Kinases; C025630:CAGCTG-specific type II deoxyribonucleases; D015252:Deoxyribonucleases, Type II Site-Specific; C024405:GATC-specific type II deoxyribonucleases