| 摘要 |
<p>The disclosure provides a method of sequencing a nucleic acid molecule that contains two or more target regions to be sequenced (such as, for example, barcodes). The invention is advantageous for sequencing by synthesis two or more target regions whose combined lengths plus the length of any intermediate sequence exceeds the available read length on a given sequencing platform. The methods of the invention utilize nucleic acid constructs containing at least the following elements: a complement of a first universal primer, a first target sequence, an optional polynucleotide spacer, a complement of a second universal primer, and a second target sequence. Both universal primers are hybridized to the template nucleic acids, however, one of the universal primers is reversibly blocked. A first round of sequencing by synthesis is performed to sequence by elongating the unblocked universal primer. Once the sequence of a target region is obtained, the first round of sequencing is terminated. Thereafter, the remaining primer is deblocked, and a second round of sequencing by synthesis is conducted to sequence a second target region.</p> |
