Highly alkaline electrolyte for single-stranded DNA separations by electrophoresis in bare silica capillaries.
A new, highly denaturing electrolyte system based on a solution containing 0.01 M NaOH, 0.0015 M Na2B4O5(OH)4 and a replaceable polymer sieving medium was designed for the separation of single-stranded DNA fragments in bare fused-silica capillaries. Extreme denaturing power, together with the optimized composition of the electrolyte, allows for a separation efficiency as high as 2,300,000 height equivalents to a theoretical plate per meter. Sample denaturation in alkaline solutions provides single-stranded DNA fragments without any intra- or intermolecular interactions at room temperature. Their electrophoretic mobilities were found to be twice those of fragments denatured by dimethylformamide or HCl. This can be interpreted in terms of an increased effective charge on the DNA molecules. The surprisingly weak electroosmosis (6 x 10(-10) m2 V-1 s-1) of polymer solutions at pH 12 or higher is considered to be the result of the dissolution of the silica capillary wall. A highly viscous thin layer of dissolved silica probably causes a shift of the slipping plane further away from the wall to the lower value of the zeta potential. Applications of the electrolyte in clinical diagnostics demonstrate its remarkable properties.
Malá
Z
Z
Institute of Analytical Chemistry, Academy of Sciences of Czech Republic, Brno, Czech Republic.
Klepárník
K
K
Bocek
P
P
D004277:DNA, Single-Stranded / Q000737:chemistry / Q000302:isolation & purification*; D004573:Electrolytes; D019075:Electrophoresis, Capillary / Q000379:methods*; D009691:Nucleic Acid Denaturation; D012822:Silicon Dioxide / Q000737:chemistry; D012972:Sodium Hydroxide / Q000737:chemistry
D004277:DNA, Single-Stranded; D004573:Electrolytes; D012972:Sodium Hydroxide; D012822:Silicon Dioxide