NUCLEIC ACID SEQUENCE ANALYSIS

摘要

Provided are methods for sequencing a nucleic acid with a sequencing enzyme, e.g., a polymerase or exonuclease. The sequencing enzyme can optionally be exchanged with a second sequencing enzyme, which continues the sequencing of the nucleic acid. In certain embodiments, a template is fixed to a surface through a template localizing moiety. The template localizing moiety can optionally anneal with the nucleic acid and/or associate with the sequencing enzyme. Also provided are compositions comprising a nucleic acid and a first sequencing enzyme, which can sequence the nucleic acid and optionally exchange with a second sequencing enzyme present in the composition. Compositions in which a template localizing moiety is immobilized on a surface are provided. Also provided are methods for using data from analytical reactions wherein two different enzymes are employed, e.g., at a same or different reaction regions.

主权项

1. A method of sequencing a single template molecule, the method comprising:
localizing the single template molecule to a reaction region through association with a template localizing moiety attached to the reaction region, wherein the template localizing moiety comprises a protein lacking nascent strand synthesis activity that translocates along the single template molecule while retaining the single template molecule at the reaction region; and sequencing a polynucleotide region of the single template molecule to obtain a first polynucleotide sequence read of the polynucleotide region, wherein the polynucleotide sequence read comprises redundant sequence information for the polynucleotide region, thereby sequencing the single template molecule.