Identification of base pairs in single-nucleotide polymorphisms by MutS protein-mediated capillary electrophoresis

Analytical Chemistry
Andrei P Drabovich, Sergey N Krylov

Abstract

Single-nucleotide polymorphisms (SNPs) are widespread genomic variations, which are associated with serious health disorders and drug resistance. Multiple clinical applications and studies of global population genetics require fast and informative analysis of SNPs. Most of conventional methods sense the presence of the SNP but cannot identify the base pair in it. Here we report simple identification of base pairs in SNPs without DNA sequencing. Our approach is based on the unique ability of MutS protein to bind different single-nucleotide mismatches in DNA with different affinities. Conceptually, the DNA in question is mixed with reference DNA, melted, and reannealed. If the DNA in question has an SNP, the products of reannealing will have two different single-nucleotide mismatches, which provide a base-pair-specific signature of the SNP. The products of reannealing are mixed with MutS, equilibrated, and separated by equilibrium capillary electrophoresis of equilibrium mixtures with MutS in the run buffer. The pattern of migration times of DNAs with mismatches is used for unequivocal identification of the base pair in the SNP. In addition to its ability to identify base pairs in SNPs, the new analytical approach is fast, simple...Continue Reading

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Aug 11, 2005·Journal of the American Chemical Society·Andrei DrabovichSergey N Krylov
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Citations

Mar 25, 2014·Legal Medicine·Takashi Taki, Kazuhiko Kibayashi
Nov 29, 2014·Critical Reviews in Biotechnology·Atul Grover, P C Sharma
Sep 30, 2016·Journal of the American Chemical Society·Maria StancescuDmitry M Kolpashchikov
May 22, 2018·Nanoscale·Rebekah J KaradeemaDmitry M Kolpashchikov
Jan 25, 2007·Electrophoresis·Sergey N Krylov
Nov 28, 2007·Electrophoresis·Eun Joo SongYoung Sook Yoo
Dec 25, 2009·Analytical Chemistry·Tao LiHailin Wang
Jul 18, 2021·Physical Review. E·Anurag Upadhyaya, Sanjay Kumar

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