Method for simultaneously performing multiple amplification reactions
First Claim
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1. A method for detecting the presence of a nucleic acid in each of multiple samples, the method comprising performing within a housing of an analyzer the automated steps of:
- (a) contacting each of multiple samples containing a nucleic acid with a solid support, such that a complex comprising the nucleic acid and the solid support is formed in each of the samples, wherein the solid support comprises a magnetically-responsive particle, and wherein the complex formed in each of the samples is suspended in a fluid component of the sample;
(b) after step (a), subjecting the complex in each of the samples to a magnetic field;
(c) during step (b), purifying the nucleic acid in each of the samples by accessing and removing at least a portion of the fluid component of each of the samples from the complex;
(d) forming multiple mixtures, each of the mixtures comprising the purified nucleic acid of one of the samples of step (c) and reagents for performing a nucleic acid amplification, wherein the mixtures are separately formed in multiple receptacles of a set of receptacles;
(e) moving the set of receptacles to a temperature-controlled environment;
(f) in each of the multiple receptacles, performing nucleic acid amplification, whereby amplification products are enzymatically synthesized, each of the amplification products comprising (i) a target sequence contained in the nucleic acid or (ii) the complement of the target sequence;
(g) in each of the multiple receptacles, selectively hybridizing a detectably labeled probe to an amplification product that is one of the amplification products synthesized in step (f), thereby forming a hybrid that comprises the probe and the amplification product, wherein the amplification product comprises the target sequence or its complement, and wherein the probe hybridizes to the target sequence or its complement in the amplification product; and
(h) in each of the multiple receptacles, detecting the presence of the hybrid, wherein the presence of the hybrid in each of the multiple receptacles is an indication of the presence of the nucleic acid in each of the samples,wherein the method is performed without human intervention.
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Abstract
An automated method for performing multiple nucleic acid amplification procedures in multiple receptacles of a set of receptacles contained in an analyzer.
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Citations
30 Claims
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1. A method for detecting the presence of a nucleic acid in each of multiple samples, the method comprising performing within a housing of an analyzer the automated steps of:
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(a) contacting each of multiple samples containing a nucleic acid with a solid support, such that a complex comprising the nucleic acid and the solid support is formed in each of the samples, wherein the solid support comprises a magnetically-responsive particle, and wherein the complex formed in each of the samples is suspended in a fluid component of the sample; (b) after step (a), subjecting the complex in each of the samples to a magnetic field; (c) during step (b), purifying the nucleic acid in each of the samples by accessing and removing at least a portion of the fluid component of each of the samples from the complex; (d) forming multiple mixtures, each of the mixtures comprising the purified nucleic acid of one of the samples of step (c) and reagents for performing a nucleic acid amplification, wherein the mixtures are separately formed in multiple receptacles of a set of receptacles; (e) moving the set of receptacles to a temperature-controlled environment; (f) in each of the multiple receptacles, performing nucleic acid amplification, whereby amplification products are enzymatically synthesized, each of the amplification products comprising (i) a target sequence contained in the nucleic acid or (ii) the complement of the target sequence; (g) in each of the multiple receptacles, selectively hybridizing a detectably labeled probe to an amplification product that is one of the amplification products synthesized in step (f), thereby forming a hybrid that comprises the probe and the amplification product, wherein the amplification product comprises the target sequence or its complement, and wherein the probe hybridizes to the target sequence or its complement in the amplification product; and (h) in each of the multiple receptacles, detecting the presence of the hybrid, wherein the presence of the hybrid in each of the multiple receptacles is an indication of the presence of the nucleic acid in each of the samples, wherein the method is performed without human intervention. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14)
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15. A method for detecting the presence of a nucleic acid in each of multiple samples, the method comprising performing within a housing of an analyzer the automated steps of:
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(a) contacting each of multiple samples containing a nucleic acid with a solid support, such that a complex comprising the nucleic acid and the solid support is formed in each of the samples, wherein the solid support comprises a magnetically-responsive particle, and wherein the complex formed in each of the samples is suspended in a fluid component of the sample; (b) after step (a), subjecting the complex in each of the samples to a magnetic field; (c) during step (b), purifying the nucleic acid in each of the samples by contacting the fluid component of each of the samples with a fluid aspirator and aspirating at least a portion of the fluid component from the complex; (d) forming multiple mixtures, each of the mixtures comprising the purified nucleic acid of one of the samples of step (c) and reagents for performing a nucleic acid amplification, wherein the mixtures are separately formed in multiple receptacles of a set of receptacles; (e) moving the set of receptacles to a temperature-controlled environment; (f) in each of the multiple receptacles, performing nucleic acid amplification, amplification products are enzymatically synthesized, each of the amplification products comprising (i) a target sequence contained in the nucleic acid or (ii) the complement of the target sequence; (g) in each of the multiple receptacles, selectively hybridizing a detectably labeled probe to an amplification product that is one of the amplification products synthesized in step (f), thereby forming a hybrid that comprises the probe and the amplification product, wherein the amplification product comprises the target sequence or its complement, wherein the probe hybridizes to the target sequence or its complement in the amplification product, and wherein the hybrid is formed in solution; and (h) in each of the multiple receptacles, detecting the presence of the hybrid while the hybrid is in solution, wherein the presence of the hybrid in each of the multiple receptacles is an indication of the presence of the nucleic acid in each of the samples, wherein the method is performed without human intervention. - View Dependent Claims (16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30)
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Specification