Methods and kits for reducing non-specific nucleic acid amplification
AssigneeGeneral Electric Company
InventorsJohn Richard Nelson, Robert Scott Duthie, Sonali Jagdish Shah, Clifford Leslie Smith
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Abstract
Methods and kits for efficient amplification of nucleic acids are provided. The methods comprise in-vitro amplification of a nucleic acid template employing partially constrained primers having terminal mismatch primer-dimer structure. The methods also comprise in-vitro amplification of a nucleic acid template employing partially constrained primers having nucleotide analogues. The methods enhance efficiency of nucleic acid amplification reaction by reducing non-specific amplification reactions.
Claims20
A kit for nucleic acid amplification, comprising:
- a nucleic acid polymerase;
- and
- a primer solution,
- wherein the primer solution consists essentially of a partially constrained primer having a terminal mismatch primer-dimer structure, and wherein the partially constrained primer consists of a nucleotide sequence (W)x(N)y(S)z, wherein x, y and z are integer values independent of each other, and wherein the value of x is 2 or 3, the value of y is 2, 3 or 4, and the value of z is 1 or 2.
A kit for isothermal nucleic acid amplification, comprising:
- a nucleic acid polymerase;
- and
- a primer solution,
- wherein the primer solution consists essentially of a partially constrained primer mixture, wherein the partially constrained primer mixture comprises a nucleotide analogue, and wherein the partially constrained primer mixture consists of a nucleotide sequence (W)X(N)y(S)Z, wherein x, y and z are integer values independent of each other, and wherein the value of x is 2 or 3, the value of y is 2, 3 or 4, and the value of z is 1 or 2.
A kit for isothermal nucleic acid amplification, comprising:
- a nucleic acid polymerase;
- and
- a primer solution,
- wherein the primer solution consists essentially of a nuclease-resistant, partially constrained primer mixture, wherein the nuclease-resistant partially constrained primer mixture comprises a modified nucleotide, wherein the nuclease-resistant, partially constrained primer has a terminal primer-dimer structure, and wherein the partially constrained primer mixture consists of a nucleotide sequence (W)x(N)y(S)z, wherein x, y and z are integer values independent of each other, and wherein the value of x is 2 or 3, the value of y is 2, 3 or 4, and the value of z is 1 or 2.
Dependencies are inferred from the claim text.