Sequencing of Small DNA Fragments with Aggregated-Induced-Emission Molecule-Labeled Nucleotides.

Feifei Sun, Shengnan Zhao, Manshu Peng, Qiang Fu, Huimin Gao, Yijing Jia, Na Na, Jin Ouyang

Analytical chemistry 2020 May 19

filter terms:

Sequencing by synthesis is a significant method for high-throughput DNA sequencing. Herein, we synthesized terminal aggregated-induced-emission luminogen (AIEgen) labeled nucleotides (dNTPs-HCAP) that could serve as substrates for some polymerases and applied them into the sequencing of small DNA fragments. In the process of DNA amplification, ratiometric AIEgens are released from dNTPs-HCAP and aggregate through the effects of phosphatase, which results in changes in the ratiometric fluorescent signals. With the AIEgen-labeled nucleotides, we accomplished the sequencing of small DNA fragments through double changes in fluorescence. In addition, we achieved the differentiation of single nucleotide polymorphisms through rolling circle amplification reactions without the addition of signal probes, which is fast and cost-effective. The introduction of ratiometric AIEgens into DNA synthesis makes the detection of DNA sequences more efficient and accurate. Therefore, the development of AIEgen-labeled nucleotides is meaningful for the study of DNA sequencing methods.

Citation

Feifei Sun, Shengnan Zhao, Manshu Peng, Qiang Fu, Huimin Gao, Yijing Jia, Na Na, Jin Ouyang. Sequencing of Small DNA Fragments with Aggregated-Induced-Emission Molecule-Labeled Nucleotides. Analytical chemistry. 2020 May 19;92(10):7179-7185