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US 2025/0019752 A1Published application
Compositions and methods for reducing master mix contamination
AssigneeLife Technologies Corporation
InventorsDaniel Philippe Blanchard, Maria Cinta Gomez Silvan
Life of this patent
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Abstract
This disclosure describes master mix compositions, nucleic acid amplification kits, methods of manufacturing master mix compositions, and methods of using master mix compositions that minimize or eliminate the negative effects of contaminant nucleic acids. Conventional master mix compositions often include contaminant nucleic acids.
Claims77
A composition comprising:
- a DNA polymerase;
- and
- an intercalating agent having the formula
- X—Ar—R1
- wherein
- X comprises an azide, a charged N, an amine, or an amino, wherein when X comprises an amino/amine, the amino/amine is NH2, NR2H, or NR2R3, wherein R2 and R3 are independently a DNA molecule to which the intercalating agent is covalently bonded or a group comprising an alkyl,
- Ar is a heterocyclic aromatic group with a ring structure having at least one charged N, and
- R1 is attached to the charged N of the Ar and comprises an alkyl.
5 more elements
A method of manufacturing a composition for use in amplification of nucleic acid, the method comprising:
- providing a first mixture that comprises one or more nucleic acid amplification components;
- adding to the first mixture an intercalating agent having the formula:
- X—Ar—R1
- wherein
- X comprises an azide, a charged N, an amine, or an amino,
- Ar is a heterocyclic aromatic group with a ring structure having at least one charged N, and
- R1 is attached to the charged N of the Ar and comprises an alkyl;
- and
- exposing the first mixture to light to activate the intercalating agent and cause covalent bonding between the intercalating agent and contaminant DNA present in the first mixture.
6 more elements
A method of manufacturing a composition for use in amplification of nucleic acid, the method comprising:
- providing a first mixture that comprises one or more nucleic acid amplification components;
- adding to the first mixture an intercalating agent having the formula:
- X—Ar—R1
- wherein
- X comprises an azide, a charged N, an amine, or an amino,
- Ar is a heterocyclic aromatic group with a ring structure having at least one charged N, and
- R1 is attached to the charged N of the Ar and comprises an alkyl and optionally a charged N;
- exposing the first mixture to light to activate the intercalating agent and cause covalent bonding between the intercalating agent and contaminant DNA present in the first mixture;
- providing a second mixture that comprises one or more nucleic acid amplification components;
- autoclaving the second mixture for at least 30 minutes;
- and
- combining the first mixture and the second mixture to form a combined mixture.
9 more elements
Dependencies are inferred from the claim text.