Compositions and methods for enhancing polynucleotide amplification reactions
AssigneeThe Trustees of Princeton University
InventorsRaj Chakrabarti, Clarence Schutt
Life of this patent
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Abstract
Compositions and methods for enhancing PCR and other polynucleotide replication reactions are disclosed. These comprise the addition of low molecular weight organic amides, sulfones or sulfoxides to PCR or other replication reaction mixtures.
Claims13
1-3. (canceled)
A composition for performing a polynucleotide replication reaction, which comprises a buffer, one or more template polynucleotides, nucleotide triphosphates, one or more polymerase enzymes or fragments thereof, and one or more reaction adjuvants comprising compounds of the formula:
- wherein:
- R1 is C or S, and
- when R1 is C, X is ═O, R3 is N and R is absent;
- when R1 is S, X is ═O or
- and R3 is C;
- R2 is H or CH3 only when one or more of R4, R5 and R6 is not H, and otherwise R2 is an unsubstituted or halogen-, hydroxyl- or alkoxy-substituted alkyl or cycloalkyl of length m, wherein m is selected such that the total number of carbons in the compound is between 3 and 8 when R1 is C and between 2 and 8 when R1 is S:
- wherein any two of R2, R3, R4, R5 and R6 can form a cyclic structure in which cyclization is effected through a bond between them;
- R4, R5 and R6 each is H, alkyl, cycloalkyl, halogen-, hydroxyl or alkoxy-substituted alkyl or cycloalkyl of length n, wherein n is selected such that the total number of carbons in the compound is between 3 and 8 when R1 is C and between 2 and 8 when R1 is S;
- when R4 and R5 are CH3, R2 cannot be H or CH3;
- wherein the reaction adjuvant comprises a cyclic compound, wherein the cyclization is effected through a bond between any two of R2, R3, R4, R5, and R6;
- wherein the cyclic portion of the compound comprises five, six or seven members:
- and
- wherein R1 is C, R3 is N, R6 is absent and of R4 or R5 is H.
(canceled)
A composition for performing a polynucleotide replication reaction, which comprises a buffer, one or more template polynucleotides, nucleotide triphosphates, one or more polymerase enzymes or fragments thereof, and one or more reaction adjuvants comprising compounds of the formula:
- wherein:
- R1 is C or S;
- and
- when R1 is C, X is ═O, R3 is N and R6 is absent;
- when R1 is S, X is ═O or
- and R3 is C:
- R2 is H or CH3 only when one or more of R4, R5and R6 is not H, and otherwise R2 is an unsubstituted or halogen-, hydroxyl- or alkoxy-substituted alkyl or cycloalkyl of length m, wherein m is selected such that the total number of carbons in the compound is between 3 and 8 when R1 is C and between 2 and 8 when R1 is S;
- wherein any two of R2, R3, R4, R5 and R6 can form a cyclic structure in which cyclization is effected through a bond between them;
- R4, R5 and R6 each is H, alkyl, cycloalkyl, halogen-, hydroxyl or alkoxy-substituted alkyl or cycloalkyl of length n, wherein n is selected such that the total number of carbons in the compound is between 3 and 8 when R1 is C and between 2 and 8 when R1 is S;
- when R4 and R5 are CH3, R2 cannot be H or CH3;
- wherein the reaction adjuvant comprises a compound in which R1 is C, X is ═O, R3 is N and R6 is absent.
15-70. (canceled)
Dependencies are inferred from the claim text.