METHOD OF SEQUENCING NUCLEIC ACID WITH UNNATURAL BASE PAIRS

摘要

Disclosed is a method of sequencing a nucleic acid containing an unnatural base pair (UBP), comprising performing two or more replacement replication reactions wherein the nucleic acid is replicated using two or more intermediate of the unnatural base pair; sequencing the nucleic acid resulting from the replacement replication reactions; clustering the sequenced nucleic acid and identifying a candidate position of the unnatural base pair; determining a ratio of conversion of the intermediate to each one of a natural base pair at the candidate position of the unnatural base pair; comparing the ratio of conversion of the intermediate to a library of pre-determined conversion rate based on the sequences of one or more natural base pair adjacent to the candidate position of the unnatural base pair; wherein a substantial match of the ratio of conversion of the intermediate to a value in the library of the pre-determined conversion rate confirms the position of the unnatural base pair, thereby determining the sequence of the nucleic acid containing the unnatural base pair. Also disclosed is an apparatus for performing the method as disclosed herein.