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Related Experiment Videos

Reversible thermo-responsive sieving matrix for oligonucleotide separation.

Jun Zhang1, Marcus Gassmann, Weidong He

  • 1Department of Chemistry, Stony Brook University, Stony Brook, NY, 11794-3400, USA.

Lab on a Chip
|March 31, 2006
PubMed
Summary
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Researchers developed a novel thermo-responsive Pluronic copolymer gel for oligonucleotide separation using microchip capillary electrophoresis. This system enables rapid and precise separation of DNA fragments within seconds.

Area of Science:

  • Polymer Chemistry
  • Analytical Chemistry
  • Biotechnology

Background:

  • Pluronic copolymers F87 and F127 exhibit temperature-dependent viscosity and dynamic coating properties in aqueous solutions.
  • These triblock copolymers (oxyethylene-oxypropylene-oxyethylene) offer tunable sol-gel transition temperatures.
  • Oligonucleotide separation is crucial in molecular biology and diagnostics.

Purpose of the Study:

  • To develop and characterize a reversible thermo-responsive gel system for oligonucleotide separation.
  • To investigate the use of Pluronic copolymer mixtures in microchip capillary electrophoresis.
  • To optimize the gel composition and transition temperature for efficient DNA fragment separation.

Main Methods:

  • Utilized a mixture of Pluronic F87 and F127 copolymers at approximately 30% (w/v) concentration.

Related Experiment Videos

  • Adjusted the F87/F127 weight ratio to control the sol-gel transition temperature between 17°C and 38°C.
  • Employed microchip-based capillary electrophoresis with a 1.5 cm separation channel.
  • Validated the sieving ability using oligonucleotide sizing markers (8-32 bases) and an Agilent Bioanalyzer 2100.

    • Main Results:

    • Achieved successful separation of oligonucleotide sizing markers ranging from 8 to 32 bases.
    • Identified an optimal F87/F127 weight ratio of 1:2, yielding a sol-gel transition at ~26°C with superior sieving ability.
    • Demonstrated fast separations within 40 seconds with single-base resolution.
    • Confirmed the gel's sieving performance using a commercial automated electrophoresis system.

    Conclusions:

    • A reversible thermo-responsive Pluronic copolymer gel system is effective for microchip capillary electrophoresis-based oligonucleotide separation.
    • The tunable sol-gel transition temperature and sieving ability of the Pluronic mixture allow for precise control over separation parameters.
    • This novel approach offers a fast, efficient, and high-resolution method for analyzing oligonucleotide fragments.