Abstract:
PROBLEM TO BE SOLVED: To provide systems, methods, and an apparatus for closed multi-stage nucleic acid amplification reactions wherein a portion of a prior-stage reaction mixture serves as the sample for the next stage reaction.SOLUTION: The invention is directed to systems, methods and an apparatus for carrying out multi-stage amplification reactions, especially under fluidly closed conditions. In one aspect, the methods of the invention are carried out in a fluidly closed reaction system that permits the isolation of a portion of a first (or prior) reaction mixture and its use as a sample or specimen in a second (or subsequent) reaction mixture, thereby substantially avoiding interfering effects that first reaction components may have in the second reaction if both reaction mixtures were simply combined together. In the aspect, the systems, methods, and apparatus of the invention may be used with any amplification reaction that permits multiple stages of amplification based on the use of nested primers.
Abstract:
PROBLEM TO BE SOLVED: To provide systems, methods, and apparatus for closed multistage nucleic acid amplification reactions wherein a part of a prior-stage reaction mixture serves as the sample for the next stage reaction.SOLUTION: There are provided systems, methods, and apparatus for carrying out multistage amplification reactions, especially under fluidly closed conditions. In one embodiment are carried out in a fluidly closed reaction system that permits the isolation of a part of a first (or prior) reaction mixture and its use as a sample or specimen in a second (or subsequent) reaction mixture, thereby substantially avoiding interfering effects that first reaction components may have in the second reaction if both reaction mixtures were simply combined together. In the above embodiment, the systems, the methods, and the apparatus may be used with any amplification reaction that permits multiple stages of amplification based on the use of nested primers.
Abstract:
Un método para realizar una reacción de amplificación anidada, comprendiendo el método las etapas de: a) realizar una reacción de amplificación de primer estadio en una cámara de reacción de un sistema cerrado de forma fluida, comprendiendo la reacción de amplificación de primer estadio uno o más polinucleótidos diana de una muestra usando reactivos de amplificación de primer estadio en una primera mezcla de reacción para formar uno o más primeros amplicones, incluyendo los reactivos de amplificación de primer estadio cebadores iniciales para cada polinucleótido diana, en donde el o los primeros amplicones se amplifican en presencia de un indicador fluorescente capaz de generar una señal óptica relacionada con una cantidad de un amplicón en la reacción de amplificación de primer estadio; b) controlar la señal óptica del indicador fluorescente en la reacción de amplificación de primer estadio en la cámara de reacción; c) retener automáticamente una parte efectiva de la mezcla de reacción de la reacción de amplificación de primer estadio cuando la señal óptica alcanza o supera un nivel umbral; d) amplificar en la cámara de reacción el o los primeros amplicones en la parte efectiva usando reactivos de amplificación de segundo estadio en una segunda mezcla de reacción para formar uno o más segundos amplicones, incluyendo los reactivos de amplificación de segundo estadio al menos un cebador secundario para cada uno del o los primeros amplicones, de modo que cada cebador secundario esté anidado en dicho primer amplicón en relación con un cebador inicial de dicho primer amplicón; y e) detectar el o los segundos amplicones para determinar la presencia o ausencia del o los polinucleótidos diana en la muestra.
Abstract:
The invention is directed to systems, methods and apparatus for carrying out multi-stage amplification reactions, especially under fluidly closed conditions. In one aspect, methods of the invention are carried out in a fluidly closed reaction system that permits the isolation of a portion of a first (or prior) reaction mixture and its use a s a sample or specimen in a second (or subsequent) reaction mixture, thereby substantially avoiding interfering effects that first reaction components may have in the second reaction if both reaction mixture were simply combined together. In the aspect, systems, methods, and apparatus of the invention may be used with any amplification reaction that permits multiple stages of amplification based on the use of nested primers.
Abstract:
The invention is directed to systems, methods and apparatus for carrying out multi-stage amplification reactions, especially under fluidly closed conditions. In one aspect, methods of the invention are carried out in a fluidly closed reaction system that permits the isolation of a portion of a first (or prior) reaction mixture and its use a s a sample or specimen in a second (or subsequent) reaction mixture, thereby substantially avoiding interfering effects that first reaction components may have in the second reaction if both reaction mixture were simply combined together. In the aspect, systems, methods, and apparatus of the invention may be used with any amplification reaction that permits multiple stages of amplification based on the use of nested primers.
Abstract:
The invention is directed to systems, methods, and apparatus for carrying out multi-stage amplification reactions, especially under fluidly closed conditions. In one aspect, methods of the invention are carried out in a fluidly closed reaction system that permits the isolation of a portion of a first (or prior) reaction mixture and its use as a sample or specimen in a second (or subsequent) reaction mixture, thereby substantially avoiding interfering effects that first reaction components may have in the second reaction if both reaction mixtures were simply combined together. In this aspect, systems, methods, and apparatus of the invention may be used with any amplification reaction that permits multiple stages of amplification based on the use of nested primers.
Abstract:
The invention is directed to systems, methods, and apparatus for carrying out multi-stage amplification reactions, especially under fluidly closed conditions. In one aspect, methods of the invention are carried out in a fluidly closed reaction system that permits the isolation of a portion of a first (or prior) reaction mixture and its use as a sample or specimen in a second (or subsequent) reaction mixture, thereby substantially avoiding interfering effects that first reaction components may have in the second reaction if both reaction mixtures were simply combined together. In this aspect, systems, methods, and apparatus of the invention may be used with any amplification reaction that permits multiple stages of amplification based on the use of nested primers.
Abstract:
The invention is directed to systems, methods, and apparatus for carrying out multi-stage amplification reactions, especially under fluidly closed conditions. In one aspect, methods of the invention are carried out in a fluidly closed reaction system that permits the isolation of a portion of a first (or prior) reaction mixture and its use as a sample or specimen in a second (or subsequent) reaction mixture, thereby substantially avoiding interfering effects that first reaction components may have in the second reaction if both reaction mixtures were simply combined together. In this aspect, systems, methods, and apparatus of the invention may be used with any amplification reaction that permits multiple stages of amplification based on the use of nested primers.
Abstract:
The invention is directed to systems, methods, and apparatus for carrying out multi-stage amplification reactions, especially under fluidly closed conditions. In one aspect, methods of the invention are carried out in a fluidly closed reaction system that permits the isolation of a portion of a first (or prior) reaction mixture and its use as a sample or specimen in a second (or subsequent) reaction mixture, thereby substantially avoiding interfering effects that first reaction components may have in the second reaction if both reaction mixtures were simply combined together. In this aspect, systems, methods, and apparatus of the invention may be used with any amplification reaction that permits multiple stages of amplification based on the use of nested primers.