METHODS FOR SEQUENCING A BIOMOLECULE BY DETECTING RELATIVE POSITIONS OF HYBRIDIZED PROBES

摘要

A sequencing method is presented in which a biomolecule is hybridized with a specially chosen pool of different probes of known sequence which can be electrically distinguished. The different probe types are tagged such that they can be distinguished from each other in a Hybridization Assisted Nanopore Sequencing (HANS) detection system, and their relative positions on the biomolecule can be determined as the biomolecule passes through a pore or channel. The methods eliminate, resolve, or greatly reduce ambiguities encountered in previous sequencing methods.

主权项

1. A method for determining a sequence of a biomolecule, the method comprising the steps of:
(a) identifying a set of k-1-length subsequences that represent a plurality of substrings of a sequence string s of the biomolecule; (b) for each of the k-1-length subsequences, identifying a pool of probes comprising four different k-mer extensions of the k-1-length subsequence; (c) for each pool identified in step (b):
(i) hybridizing the biomolecule with the four k-mer probes making up the pool; and(ii) detecting relative positions of the k-mer probes that have attached to the biomolecule; and (d) ordering the subsequences corresponding to the detected attached probes to determine the sequence string s of the biomolecule.