The use of poly(2-acrylamido-2-methyl-1-propanesulfonic acid) polymers as spacers for isotachophoresis in sieving gel matrices.
The electric field strength gradients generated in isotachophoresis (ITP) may be used for the separation of biomolecules. Poly(2-acrylamido-2-methyl-1-propanesulfonic acid) (polyAMPS) polymers of a uniform distribution of molecular mass were synthesized and used as novel spacers in ITP. Since these polymeric spacers are strongly acidic species, their ionic charges remain constant over a wide pH range, so that their ionic mobilities are governed solely by their molecular masses and not by the pH of the milieu. A modification of ITP known as telescope electrophoresis was used to separate a number of acidic dyes of varying ionic mobility, using polyAMPS polymers as spacers. The resolution obtained was superior to that obtained by polyacrylamide gel electrophoresis (PAGE), due to the focusing effect of the electric field strength gradient. Since these novel polymeric spacers are designed to operate within sieving medium, it was decided to test their suitability for the separation of DNA molecules. DNA molecules up to 1000 bp long were successfully resolved, with a similar resolution to that obtained with conventional PAGE.
Department of Biochemistry, University of the Witwatersrand, Johannesburg, South Africa.
M P
MP
Bellini
K L
KL
Manchester
D004396:Coloring Agents / Q000737:chemistry; D004247:DNA / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D011108:Polymers / Q000737:chemistry*; D013451:Sulfonic Acids / Q000737:chemistry*; D013864:Thioglycolates / Q000737:chemistry; D014783:Viscosity
D004396:Coloring Agents; D011108:Polymers; D013451:Sulfonic Acids; D013864:Thioglycolates; C077118:poly(2-acrylamido-2-methyl-1-propanesulfonic acid); D004247:DNA