CPC G16B 30/00 (2019.02) [C12Q 1/6806 (2013.01); C12Q 1/689 (2013.01); C12Q 1/6855 (2013.01); C12Q 1/6869 (2013.01); G16B 30/10 (2019.02)] | 13 Claims |
1. A method of obtaining nucleotide sequence information of a double-stranded portion of a circular nucleic acid, the method comprising:
a) providing a circular nucleic acid comprising a double-stranded portion, the double-stranded portion comprising a sense strand and an antisense strand of said sense strand complementary to the sense strand, wherein the sense strand and the antisense strand of the double-stranded portion are linked at their both ends to form said circular nucleic acid,
b) performing a rolling circle replication reaction using said circular nucleic acid as a replication template, thereby producing a concatemer having segments comprising repeating copies of said sense strand and said antisense strand;
c) performing a single-molecule nucleic acid sequencing process on the concatemer produced in step b), thereby generating a sequencing read from said concatemer, said sequencing read comprising multiple reads of at least one first sequence corresponding to said sense strand and multiple reads of at least one second sequence corresponding to said antisense strand; and
d) comparing or assembling said multiple reads of at least one first sequence and said multiple reads of at least one second sequence from said sequencing read, thereby obtaining the nucleotide sequence information of said double-stranded portion of said circular nucleic acid.
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