Patent ID: 11891659
Assignee: PACIFIC BIOSCIENCES OF CALIFORNIA, INC.
Field: Biotechnology (Chemistry)
Classification: CPC C | IPC C

Claim 16:
17. A method of sequencing a DNA template, the method comprising:
a) providing a reaction mixture comprising:
the DNA template,
a replication initiating moiety that complexes with or is integral to the template,
at least one nucleotide analog, which analog comprises a polypeptide moiety positioned between a dye moiety and a nucleotide component of the analog, wherein the polypeptide moiety comprises at least 60 amino acids; and
a recombinant DNA polymerase, which recombinant polymerase comprises an amino acid sequence that is at least 80% identical to SEQ ID NO:1 or at least 80% identical to SEQ ID NO:2, and which recombinant polymerase comprises an amino acid substitution at position D510, wherein identification of positions is relative to SEQ ID NO:1, wherein the recombinant polymerase is capable of replicating at least a portion of the template using the replication initiating moiety in a template-dependent polymerization reaction;

b) subjecting the reaction mixture to a polymerization reaction in which the recombinant polymerase replicates at least a portion of the template in a template-dependent manner, whereby the at least one nucleotide analog is incorporated into the resulting DNA; and
c) identifying a time sequence of incorporation of the at least one nucleotide analog into the resulting DNA.