ENZYMES RESISTANT TO PHOTODAMAGE

摘要

Provided are compositions comprising modified DNA polymerases that exhibit improved photostability compared to the parental polymerases from which they were derived. Provided are methods for generating enzymes, such as DNA polymerases, with the aforementioned phenotype. Provided are methods of using polymerases with increased resistance to photodamage to make a DNA or to sequence a DNA template.

主权项

1. A composition comprising:
a triplet state quencher, a fluorescently labeled or fluorogenic nucleotide analog, and a recombinant DNA polymerase to which the triplet state quencher is covalently or noncovalently bound, wherein the nucleotide analog serves as a substrate for the polymerase, wherein the nucleotide analog does not comprise the triplet state quencher, and wherein the triplet state quencher serves as an acceptor for the excited triplet state of the fluorescently labeled analog or of the fluorogenic analog's product.