IDENTIFYING MODIFIED BASES USING HEMI-NATURAL NUCLEIC ACIDS

摘要

Methods, compositions, and systems are provided for characterization of modified nucleic acids. Methods are provided for sequencing hemi-natural nucleic acids such as hemi-genomic DNA, having two complementary strands, one a natural sequence and the other a synthetic sequence. The identification of modified bases can be enhanced by comparing the sequencing information from the natural sequence, which has, for example, natural base modifications, with the synthetic sequence, which typically has no base modifications. The presence and identity of a modified base can be determined by monitoring kinetics, for example the kinetics of polymer meditated nucleic acid synthesis.

主权项

1. A method for single molecule sequencing and base modification detection comprising:
providing a template nucleic acid comprising a double-stranded portion and at least one hairpin loop connecting the two complementary strands, wherein one strand of the double stranded portion comprises a natural sequence, and the complementary strand comprises a synthetic sequence; carrying out single molecule sequencing to obtain sequencing information whereby the sequencing information from the strand comprising the natural sequence is compared with the sequencing information of the strand comprising the synthetic sequence to determine the presence of modified bases on the natural nucleic acids.