Mutant Taq polymerase resistant to inhibition of amplification in the presence of cyanine dye
The invention includes a mutant Taq polymerase or biologically active fragment thereof, which can significantly extend and amplify a target sequence in comparison to wild type Taq polymerase, where the extension mixture includes asymmetrical cyanine dye, including SYBR Green I. The mutant Taq polymerase, or a biologically active fragment thereof, has one or more substitutions differing from the wild type as shown in Table I.
1. A composition comprising a mutant Taq polymerase having at least 99% amino acid sequence identity to the sequence of wild type Taq polymerase as shown in SEQ ID NO: 484, wherein said mutant Taq polymerase has the amino acid mutation R801 D, wherein said composition includes a cyanine dye and said mutant Taq polymerase is resistant to inhibition of amplification by the cyanine dye.
2. The composition of claim 1 wherein the cyanine dye is an asymmetrical cyanine dye.
3. The composition of claim 2 wherein the cyanine dye is SYBR Green 1.
4. The composition of claim 1 wherein the mutant Taq polymerase has more than 99% amino acid sequence identity to the sequence of wild type Taq polymerase as shown in SEQ ID NO: 484.