| 摘要 |
<p>A decoding and sequencing method by real-time synthesis of two nucleotides into DNA. Single-sequencing reactions are performed on two different nucleotides, namely, X and Y simultaneously, and a base sequence fragment code XYn is obtained according to the quantitative relation between the number of synthesizing nucleotides and the number of the detected molecules which are generated in real time. The entire sequencing comprises performing two sets of sequencing reactions on the same template, wherein each set of sequencing comprises four nucleotides, namely, dATP, dCTP, dGTP, and dTTP, and cycles of sequencing reactions by simultaneous synthesis of two different nucleotides are performed in a manner that each nucleotide is used only once in each cycle. After several sequencing reactions, a set of XYn information ranked in a sequencing order is obtained. When said set of sequencing reactions is completed, denaturation is performed to eliminate an extended strand of a sequencing primer and the sequencing primer is re-crossed to perform a second set of sequencing reactions, and then several XYn information ranked by the second sequencing reaction is obtained; and finally, a specific base sequence of a nucleic acid fragment to be detected is assembled by decoding two sets of XYn information ranked in a sequencing order.</p> |
