CUANTIFICACION DE ACIDOS NUCLEICOS UTILIZANDO LA REACCION EN CADENA DE LA POLIMERASA

• Main Authors: MARK, DAVID, F, WANG, ALICE, M, DOYLE, MICHAEL, V
• Format: Patent
• Language: Spanish
• Published: 16.08.1999
• Edition: 6
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; COMPOSITIONS THEREOF; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; CULTURE MEDIA; ENZYMOLOGY; INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIRCHEMICAL OR PHYSICAL PROPERTIES; MEASURING; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; PHYSICS; PROCESSES OF PREPARING SUCH COMPOSITIONS; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; TESTING; VINEGAR; WINE

LA PRESENTE INVENCION PRESENTA UN METODO PARA DETERMINAR LA CANTIDAD DE UN SEGMENTO DE ACIDO DEL BLANCO EN UNA MUESTRA MEDIANTE LA REACCION EN CADENA DE LA POLIMERASA. EL METODO CONSISTE EN LA AMPLIFICACION SIMULTANEA DEL SEGMENTO DE ACIDO NUCLEICO DEL BLANCO Y DE UN SEGMENTO DE ACIDO NUCLEICO ESTANDAR INTERNO. LA CANTIDAD DE ADN AMPLIFICADO DE CADA SEGMENTO SE DETERMINA Y COMPARA CON LAS CURVAS ESTANDAR PARA DETERMINAR LA CANTIDAD DEL SEGMENTO DE ACIDO NUCLEICO DEL BLANCO PRESENTE EN LA MUESTRA ANTES DE LA AMPLIFICACION. EL METODO ES ESPECIALMENTE PREFERENTE PARA DETERMINAR LA CANTIDAD DE UNA ESPECIE DE MARN ESPECIFICA EN UNA MUESTRA BIOLOGICA. ADICIONALMENTE, SE PRESENTA UN ESTANDAR INTERNO UTIL PARA LA CUANTIFICACION DE MULTIPLES ESPECIES DE MARN. The present invention provides a method for determining the amount of a target acid segment in a sample by polymerase chain reaction. The method involves the simultaneous amplification or the target nucleic acid segment and an internal standard nucleic acid segment. The amount of amplified DNA from each segment is determined and compared to standard curves to determine the amount of the target nucleic acid segment present in the sample prior to amplification. The method is especially preferred for determining the quantity of a specific mRNA species in a biological sample. Additionally, an internal standard is provided useful for quantitation of multiple mRNA species.