Patent ID: 11891651
Assignee: ELEMENT BIOSCIENCES, INC.
Field: Measurement (Instruments)
Classification: CPC C  G | IPC C  G

Claim 27:
28. The method of claim 1, wherein the forward sequencing of step (b) and the reverse sequencing of step (e) comprise:
a) contacting a plurality of a first polymerases with (i) a plurality of nucleic acid template molecules and (ii) a plurality of soluble sequencing primers, wherein the contacting is conducted under conditions suitable to form a plurality of first complexed polymerases comprising a first sequencing polymerase bound to nucleic acid duplexes, wherein the nucleic acid duplexes comprise the nucleic acid template molecule hybridized to the sequencing primer, wherein (1) the plurality of nucleic acid template molecules comprise a plurality of the immobilized concatemer template molecules and the plurality of soluble primers comprise a plurality of the soluble forward sequencing primers, or wherein (2) the plurality of nucleic acid template molecules comprise a plurality of the retained forward extension strands and the plurality of soluble sequencing primers comprise a plurality of the soluble reverse sequencing primers;
b) contacting the plurality of first complexed polymerases with a plurality of detectably labeled multivalent molecules, wherein individual multivalent molecules comprise a core attached to multiple nucleotide arms, wherein each nucleotide arm is attached to a nucleotide unit, thereby forming a plurality of multivalent-complexed polymerases, under conditions suitable for binding complementary nucleotide units to at least two of the plurality of first complexed polymerases, thereby forming a plurality of multivalent-complexed polymerases, and wherein the conditions inhibit incorporation of the complementary nucleotide units into the sequencing primers of the nucleic acid duplexes;
c) detecting the plurality of multivalent-complexed polymerases; and
d) identifying the nucleo-base of the complementary nucleotide units that are bound to the plurality of first complexed polymerases in the plurality of multivalent-complexed polymerases, thereby determining the sequence of the nucleic acid template.