Rapid identification of single nucleotide polymorphisms by fluorescence-based capillary electrophoresis.

摘要: We describe the application of two different fluores- cence-based techniques (ddNTP primer extension and single-strand con- formation polymorphism (SSCP)) to detection single nucleotide polymorphisms (SNPs) by capillary electrophoresis. The ddNTP technique is based on extension, in presence fluo- rescence-labeled dideoxy nucleotides (ddNTP, terminators), an unla- beled oligonucleotide that binds complementary template immediately adjacent mutant position. Given there are no unlabeled dNTPs, a added its 3' end, resulting fluorescence-labeled product which readily sepa- rated On other hand, non-radioiso- topic version SSCP established this study uses fluorescent dye label PCR products, also analyzed electro- phoresis. These procedures were used identify well-defined SNP exon 7 human p53 gene DNA samples isolated from hu- man cell lines (CEM THP-1 cells). results revealed heterozy- gous single-base transition (G A) at position 14071 CEM cells, proving both fluorescence-based rapid, simple, robust, specific with ambiguity inter- pretation for SNPs.