Single-stranded DNA-binding protein facilitates gel-free analysis of polymerase chain reaction products in capillary electrophoresis.
It has been recently demonstrated that single-stranded DNA-binding protein (SSB) can facilitate quantitative analyses of DNA, RNA, and proteins in gel-free capillary electrophoresis (CE). Here, we report the application of SSB-mediated gel-free CE for analyses of polymerase chain reaction (PCR) products. The unique ability of SSB to bind ssDNA but not double-stranded DNA (dsDNA) allows efficient separation of three types of DNA molecules in the PCR reaction mixture: primers, products (amplified templates), and by-products, which originate from non-specific DNA hybridization. SSB-mediated gel-free CE analysis of PCR products combines simplicity, high sensitivity, and outstanding quantitative capabilities. The ability of the method to distinguish between products and by-products makes this method an indispensable tool in preparative PCR (e.g., in the development of nucleotide aptamers).
Drabovich
Andrei
A
Department of Chemistry, York University, Toronto, Ontario M3J IP3, Canada.
Krylov
Sergey N
SN
D017931:DNA Primers; D004268:DNA-Binding Proteins / Q000737:chemistry*; D019075:Electrophoresis, Capillary / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*
D017931:DNA Primers; D004268:DNA-Binding Proteins