Multiplex polymerase chain reaction (PCR) with color-tagged module-shuffling primers for comparing gene expression levels in various cells.
Nucleic Acids Res
; 29(16): E84, 2001 Aug 15.
Article
en En
| MEDLINE
| ID: mdl-11504892
ABSTRACT
A method based on the multiplex polymerase chain reaction (PCR) and gel electrophoresis for the comparative analysis of gene expression levels was developed. Using the method many cDNA fragments from different sources can be compared simultaneously. Competitive PCR amplification of expressed genes from different sources was performed by using 'module-shuffling primers' (MPSs). The MPSs (labeled with different fluorophores) consist of sequence modules of 3 or 4 nt. The modules are arranged in different orders in each primer; therefore, the base sequences of the primers are different but their melting temperatures are identical. The genes expressed in different sources are ligated with tags complementary with the MPSs. Tag-ligated fragments are mixed in one tube and amplified at the same amplification efficiency by the MPSs. Amplified fragments are detected separately by multiple-color gel electrophoresis. This method can detect different amounts of each expressed gene, up to a difference in amounts of 30%, and its detection limit is 0.1 amol per assay.
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Reacción en Cadena de la Polimerasa
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Color
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Cartilla de ADN
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Perfilación de la Expresión Génica
Tipo de estudio:
Diagnostic_studies
Límite:
Animals
Idioma:
En
Revista:
Nucleic Acids Res
Año:
2001
Tipo del documento:
Article
País de afiliación:
Japón