Methods and compositions for multiplex amplification of nucleic acids
AssigneeAffymetrix, Inc.
InventorsHajime Matsuzaki, Eric A. Murphy
Life of this patent
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Abstract
A method is described for predetermining ratios of primer pairs present in a single reaction vessel so as to achieve approximately equimolar yield of products. The ratios are determined as a function of the length of the amplicon and the length of other amplicons being simultaneously tested. The primers may desirably be for p53 gene sequences.
Claims13
A method for combining primers for an amplification reaction of a set of multiple amplicons including an amplicon A and an amplicon L, wherein amplicon L is the longest amplicon in said set, comprising:
- calculating primer concentrations using the formula:
- CA=CL(LA÷LL)2
- wherein CA is the concentration of primers for an amplicon A;
- wherein CL is the concentration of primers for the longest amplicon;
- wherein LA is the length of the amplicon A;
- and wherein LL is the length of the longest amplicon;
- combining the primers in a reaction vessel at the concentrations determined using said formula.
A kit comprising a set of primers for performing multiple polymerase chain reactions in a single vessel, comprising:
- twenty primers having sequences as shown in SEQ ID NO:
- ID NOS:
- 1-20.
Dependencies are inferred from the claim text.