REAL-TIME SEQUENCING METHODS AND SYSTEMS

摘要

The present invention is generally directed to compositions, methods, and systems for performing single-molecule, real-time analysis of a variety of different biological reactions. The ability to analyze such reactions provides an opportunity to study those reactions as well as to potentially identify factors and/or approaches for impacting such reactions, e.g., to either enhance or inhibit such reactions. In certain preferred embodiments, RNA templates are used in single-molecule real-time sequencing reactions.

主权项

1. A method for identifying a secondary structure of an RNA molecule, the method comprising:
(a) providing the RNA molecule, wherein the RNA molecule comprises a secondary structure important to the biological function of the RNA molecule; (b) binding a sequencing engine to the RNA molecule to produce a complex; (c) immobilizing the complex in an optical confinement such that the optical confinement contains only one such complex; (d) introducing a reaction mixture comprising a set of labeled nucleotides to the optical confinement; (e) processing the RNA template with the sequencing engine, thereby generating a sequence read for the RNA template; and (f) detecting an energetic profile of the sequencing engine during the processing, wherein the energetic profile is indicative of the secondary structure, thereby identifying the secondary structure in the RNA template within the sequence read.