Separation of DNA fragments and single strand conformation polymorphism analysis in bare capillaries using poly(acrylamide-dimethylacrylamide) as a separation medium.
A short chain poly(acrylamide-dimethylacrylamide) (PADMA) was synthesized in aqueous phase using isopropanol as a chain transfer agent, and was characterized according to the chemical composition and molecular mass. This polymer can form a stable dynamic coating on the inner surface of the capillary, thereby suppressing the electroosmotic flow and DNA-capillary wall interaction. The sieving medium has low viscosity and capillary filling with this medium and medium replacement were conveniently carried out by commercial capillary electrophoresis instruments. The effects of components and concentration of copolymers on the separation of DNA fragments were investigated. Highly efficient separation of DNA fragments, successful single strand conformation polymorphism (SSCP) analysis and good reproducibility of the migration time were obtained in bare capillaries using these copolymers as sieving media. Our preliminary results demonstrate that PADMA will become an alternative matrix for DNA separation by capillary electrophoresis.
Ren
J
J
College of Chemistry and Chemical Engineering, Hunan Normal University, Changsha, PR China. jicunren@hunnu.edu.cn
Fang
Z F
ZF
D000178:Acrylamides / Q000737:chemistry*; D001483:Base Sequence; D004247:DNA / Q000737:chemistry / Q000302:isolation & purification*; D017931:DNA Primers; D008970:Molecular Weight; D016133:Polymerase Chain Reaction; D018807:Polymorphism, Single-Stranded Conformational; D015203:Reproducibility of Results; D014783:Viscosity
D000178:Acrylamides; D017931:DNA Primers; C429789:poly(acrylamide-co-N,N-dimethylacrylamide); D004247:DNA