Simultaneous analysis of genes by capillary electrophoresis with a laser-induced fluorescence detector using a stepwise field strength gradient.
A mixture of polymerase chain reaction (PCR) products, 100, 105, 300, 310, 485, and 500 base pair (bp) DNA fragments, was analyzed by capillary electrophoresis equipped with a laser-induced fluorescence detector (CE-LIF) using a stepwise gradient of electric field strength. The optimum condition for the analysis of PCR products was 0.5% methylcellulose and 160 V/cm from 0 to 10 min and 270 V/cm from 10 to 17 min. The length (bp) of DNA could be estimated from the relationship between the relative migration time and bp length. The relative standard deviation (R.S.D.) of DNA size (bp) was less than 3.5% and the difference from the true value was only 2.4 bp.
Sumita
C
C
Department of Chemistry, Kobe Pharmaceutical University, Japan.
Tsuhako
M
M
Baba
Y
Y
D004247:DNA / Q000032:analysis*; D019075:Electrophoresis, Capillary / Q000379:methods; D005453:Fluorescence; D005796:Genes; D007834:Lasers; D016133:Polymerase Chain Reaction / Q000379:methods; D015203:Reproducibility of Results