Ultrafast detection of microsatellite repeat polymorphism in endothelin 1 gene by electrophoresis in short capillaries.
The methodology and instrumentation for fast denaturing electrophoresis in short capillaries was developed and exemplified by detection of short tandem repeat polymorphism in the endothelin 1 gene. The resolution of two nucleotides, which is required for the detection of a dinucleotide repeat polymorphism, was achieved in a capillary of an effective length of 2.5 cm at a temperature of 600C and an electric field strength of 600 V/cm in 42 s. Thus, the use of denaturing electrophoresis in short capillaries with laser-induced fluorescence detection resulted in a reduction of analysis time by a factor of 200 when compared to the conventional slab gel electrophoresis. The developed methodology and instrumentation is advantageous for an implementation in clinical diagnostics and genetic population screening where fast analytical instrumentation amenable to automation is of paramount importance.
Klepárnik
K
K
Institute of Analytical Chemistry, Academy of Sciences of the Czech Republic, Brno. klep@iach.cz
Malá
Z
Z
Pribyla
L
L
Blazková
M
M
Vasků
A
A
Bocek
P
P
D004247:DNA / Q000032:analysis* / Q000235:genetics*; D019075:Electrophoresis, Capillary / Q000379:methods*; D019332:Endothelin-1 / Q000235:genetics*; D006801:Humans; D018598:Minisatellite Repeats / Q000235:genetics*; D011110:Polymorphism, Genetic; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000379:methods
D019332:Endothelin-1; D004247:DNA