Rapid identification of single nucleotide polymorphisms by fluorescence-based capillary electrophoresis
Genet. mol. res. (Online)
;
1(1): 72-78, Mar. 2002.
Artigo
em Inglês
| LILACS
| ID: lil-417650
RESUMO
We describe the application of two different fluorescence-based techniques (ddNTP primer extension and single-strand conformation polymorphism (SSCP)) to the detection of single nucleotide polymorphisms (SNPs) by capillary electrophoresis. The ddNTP primer extension technique is based on the extension, in the presence of fluorescence-labeled dideoxy nucleotides (ddNTP, terminators), of an unlabeled oligonucleotide primer that binds to the complementary template immediately adjacent to the mutant nucleotide position. Given that there are no unlabeled dNTPs, a single ddNTP is added to its 3' end, resulting in a fluorescence-labeled primer extension product which is readily separated by capillary electrophoresis. On the other hand, the non-radioisotopic version of SSCP established in this study uses fluorescent dye to label the PCR products, which are also analyzed by capillary electrophoresis. These procedures were used to identify a well-defined SNP in exon 7 of the human p53 gene in DNA samples isolated from two human cell lines (CEM and THP-1 cells). The results revealed a heterozygous single-base transition (G to A) at nucleotide position 14071 in CEM cells, proving that both fluorescence-based ddNTP primer extension and SSCP are rapid, simple, robust, specific and with no ambiguity in interpretation for the detection of well-defined SNPs
Texto completo:
DisponíveL
Índice:
LILACS (Américas)
Assunto principal:
Leucemia Linfoide
/
Primers do DNA
/
Eletroforese Capilar
/
Polimorfismo de Nucleotídeo Único
Tipo de estudo:
Estudo diagnóstico
/
Estudo observacional
Limite:
Humanos
Idioma:
Inglês
Revista:
Genet. mol. res. (Online)
Assunto da revista:
Biologia Molecular
/
Genética
Ano de publicação:
2002
Tipo de documento:
Artigo
País de afiliação:
Espanha
Instituição/País de afiliação:
Institut d' Investigacions Biomèdiques August Pi i Sunyer/ES
Similares
MEDLINE
...
LILACS
LIS