Genotyping three human SNPs by Cy5-ddNTP primer extension / 中国药科大学学报

ABSTRACT

To establish a simple and rapid method to genotype the human single nucleotide polymorphisms(SNPs). Methods:In the presence of the appropriate Cy5-ddNTPs and dNTPs,the primer was extended by one or more bases depending on the sequence at the polymorphic site. The microchip electrophoresis was used to detect the product of extension.The switching characteristics of the primer extension reaction was increased by introducing an artificially mismatched base into the third position from the 3'-terminus of the primers. Results:Three human SNPs were detected and the alleles were distinguished on the basis of size of the extended product. All samples were accurately assayed within 100 s in microchip electrophoresis. Conclusion:This technique offers a high-speed,easy and cheap method for genotyping SNPs.