The characterization of a new size-sieving polymeric matrix for the separation of DNA fragments using capillary electrophoresis.
A low-viscosity, polysaccharide matrix (TreviSol-CE [TS-CE], Trevigen, Inc., Gaithersburg, MD, U.S.A.) has been characterized experimentally in capillary electrophoresis for the separation of DNA fragments. The mass fraction (%) of the matrix in buffer solution and the applied field strength may be varied according to the size of DNA fragments to be separated. The use of tris-phosphate-EDTA (TPE) buffer at pH 7, instead of the more commonly used tris-borate-EDTA (TBE) and tris-acetate-EDTA (TAE) buffers at pH 8.3, provides higher separation efficiency in conjunction with this matrix and simultaneously preserves the internal coating of the capillary for high run-to-run reproducibility. In comparison with other commercially available DNA separation matrices for CE, TS-CE provides enhanced separation ability of DNA fragments greater than 600 base pairs (bp) in length and lower UV absorbance background for enhanced detectability.
Siles
B A
BA
College of William and Mary, Department of Chemistry, Williamsburg, VA 23187, USA.
Collier
G B
GB
D001483:Base Sequence; D004247:DNA / Q000737:chemistry*; D019075:Electrophoresis, Capillary / Q000379:methods; D007202:Indicators and Reagents; D009838:Oligodeoxyribonucleotides / Q000737:chemistry / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods; D011134:Polysaccharides / Q000737:chemistry; D015203:Reproducibility of Results; D015183:Restriction Mapping
D007202:Indicators and Reagents; D009838:Oligodeoxyribonucleotides; D011134:Polysaccharides; D004247:DNA