Reversible thermo-responsive sieving matrix for oligonucleotide separation.
A reversible thermo-responsive gel system, consisting of Pluronic copolymer mixture of F87 and F127, has been used to successfully carry out the separation of oligonucleotides, for the first time, by microchip-based capillary electrophoresis. Pluronic triblock copolymers F87 (E(61)P(40)E(61)) and F127 (E(99)P(69)E(99)), with E, P, and subscript denoting oxyethylene, oxypropylene, and segment length respectively, have a unique temperature dependent viscosity-adjustable property and a dynamic coating ability in aqueous solution, including 1 x TBE buffer. The mixture solution has a reversible thermo-responsive property and its sol-gel transition temperature can be adjusted ranging from about 17 degrees C to 38 degrees C by varying the relative weight ratio of F87 and F127 at an optimized concentration of approximately 30% (w/v) for oligonucleotide separations. Oligonucleotide sizing markers ranging from 8 to 32 base could be successfully separated in a 1.5 cm long separation channel by the mixture solution in its gel-like state. A 30% (w/v) with a F87/F127 weight ratio of 1 ratio 2 which has a "sol-gel" transition point of about 26 degrees C shows the best sieving ability. The sieving ability of the mixture solution was further confirmed in an Agilent Bioanalyzer 2100 system. Fast separation of oligonucleotides has been achieved within 40 s with one base resolution.
Department of Chemistry, Stony Brook University, Stony Brook, NY, 11794-3400, USA.
Jun
J
Zhang
Marcus
M
Gassmann
Weidong
W
He
Fen
F
Wan
Benjamin
B
Chu
D046209:Electrophoresis, Microchip / Q000379:methods*; D006358:Hot Temperature; D008970:Molecular Weight; D009841:Oligonucleotides / Q000302:isolation & purification*; D020442:Poloxamer / Q000737:chemistry*
D009841:Oligonucleotides; D020442:Poloxamer