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US 2026/0110024 A1Published application
Pre-amplification assay
AssigneeBio-Rad Laboratories, Inc.
InventorsYanhong KONG, Steven OKINO
Life of this patent
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Abstract
Provided herein are methods and compositions to determine the efficacy of a nucleic acid pre-amplification reaction.
Claims20
A method of detecting a condition in a first amplification reaction comprising:
- assembling a first mixture, wherein the first mixture comprises a first nucleic acid template, a second nucleic acid template, a first set of primers specific for the first nucleic acid template, and a sample comprising sample nucleic acids;
- carrying out the first amplification reaction with the first mixture for a first number of cycles to generate a product;
- assembling a second mixture, wherein the second mixture comprises the product, a second set of primers specific for the second nucleic acid template, and a third set of primers specific for an amplification product of the first nucleic acid template;
- carrying out a second amplification reaction with the second mixture, wherein the second amplification reaction is quantitative;
- determining and comparing the number of cycles required to reach Cq for amplification of the first nucleic acid template (Cq1) and the number of cycles required to reach Cq for amplification of the second template (Cq2);
- and
- wherein a condition in the first amplification reaction is detected when the difference between the number of cycles required to reach Cq2 and the number of cycles required to reach Cq1 is less than a threshold number of cycles.
4 more elements
A kit comprising:
- a first container with a first set of primers specific for a first nucleic acid template;
- a second container with a second set of primers specific for a second nucleic acid template,
- the first nucleic acid template and the second nucleic acid template;
- and
- a third set of primers specific for an amplification product of the first nucleic acid template.
2 more elements
A method of detecting a condition in a first amplification reaction comprising:
- assembling a first mixture, wherein the first mixture comprises a first nucleic acid template, a second nucleic acid template, a first set of primers specific for the first nucleic acid template, and a sample comprising sample nucleic acids;
- carrying out the first amplification reaction with the first mixture for a first number of cycles to generate a product;
- assembling a second mixture, wherein the second mixture comprises the product, and reagents for carrying out a second nucleic acid assay, wherein the second nucleic acid assay is quantitative;
- carrying out the second nucleic acid assay with the second mixture;
- determining and comparing a signal indicative of the amount of an amplification product of first nucleic acid template in the second mixture and a signal indicative of the amount of second nucleic acid template in the second mixture;
- and
- wherein a condition in the first amplification reaction is detected when the difference between the amount of the amplification product of the first nucleic acid template and the amount of second nucleic acid template in the second mixture is less than a threshold value.
4 more elements
Dependencies are inferred from the claim text.