摘要 |
<p>A nucleic acid molecule to be sequenced is generated having tandem repeats of a sequence, and also having modified nucleotides which reduce the levels of secondary structure. The presence of tandemly repeated sequence and the absence of secondary structure increases the rate of sequencing and accuracy of sequences generated by nanopore sequencing. A method comprises the steps of: providing two separate, adjacent solutions of a medium and an interface between the two pools, the interface having a channel so dimensioned as to allow sequential nucleotide-by-nucleotide passage from one pool to the other pool of only one nucleic acid molecule at a time; providing a nucleic acid molecule with at least one repeat of a nucleotide sequence to be determined, wherein the nucleic acid molecule is enzymatically synthesized using a circular template, and wherein the nucleic acid molecule contains modified nucleotides that reduce secondary structure in the nucleic acid molecule; placing the nucleic acid molecule in one of the two pools; and taking measurements as each of the nucleotides of the nucleic acid molecule passes through the channel so as to determine the sequence of the nucleic acid molecule.</p> |