Bulletin of the Korean Chemical Society

Korean Chemical Society (대한화학회)
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Fast Microchip Electrophoresis Using Field Strength Gradients for Single Nucleotide Polymorphism Identification of Cattle Breeds

  • Oh, Doo-Ri (Department of Chemistry and Research Institute of Physics and Chemistry (RINPAC), Chonbuk National University) ;
  • Cheong, Il-Cheong (Hankyong National University) ;
  • Lee, Hee-Gu (Medical Genomic Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Eo, Seong-Kug (Department of Microbiology, College of Veterinary Medicine and Bio-Safety Research Institute, Chonbuk National University) ;
  • Kang, Seong-Ho (Department of Chemistry and Research Institute of Physics and Chemistry (RINPAC), Chonbuk National University)
  • Received : 2010.01.12
  • Accepted : 2010.05.05
  • Published : 2010.07.20
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Abstract

A microchip electrophoresis (ME) method was developed using a programmed field strength gradients (PFSG) for the single nucleotide polymorphism (SNP) based fast identification of cattle breeds. Four different Korean cattle (Hanwoo) and Holstein SNP markers amplified by allele-specific polymerase chain reaction were separated in a glass microchip filled with 0.5% poly(ethyleneoxide) ($M_r$ = 8 000 000) by PFSG as follows: 750 V/cm for 0 - 14 s, 166.7 V/cm for 14 - 31 s, 83.3 V/cm for 31 - 46 s, and 750 V/cm for 46 - 100 s. The cattle breeds were clearly distinguished within 45 s. The ME-PFSG method was 7 times and 5 times faster than the constant electric field ME method and the capillary electrophoresis- PFSG method, respectively, with a high resolving power ($R_s$ = 5.05 - 9.98). The proposed methodology could be a powerful tool for the fast and simultaneous determination of SNP markers for various cattle breeds with high accuracy.

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References

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