RAPID DETECTION OF COMPLEMENTARY AND MISMATCHED DNA-SEQUENCES USING FLUORESCENCE POLARIZATION

The effect of base-pair mismatches on DNA hybridization was investigat ed by fluorescence polarization to evaluate the specificity of the DNA hybridization technique. A fluorescein-labeled 24-mer DNA probe was u sed to monitor the course of hybridization, from which the rate of DNA hybridization was obtained. The specificity of the method was conside red by comparing the hybridization rate of the fluorescein-labeled pro be with complementary oligomers comprising a number of base pair misma tches. In either 0.8 M NaCl or 0.2 M MgCl2, rapid in vitro determinati on of target DNA was possible within 10 minutes if the sample DNA had 3 or less base-pair mismatches. It was shown that this method would be applicable for rapid and specific DNA determination, even in the pres ence of mismatches.