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

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 fr...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2020-05, Vol.92 (10), p.7179-7185
Main Authors: Sun, Feifei, Zhao, Shengnan, Peng, Manshu, Fu, Qiang, Gao, Huimin, Jia, Yijing, Na, Na, Ouyang, Jin
Format: Article
Language:English
Subjects:
Amplification
Chemical synthesis
Chemistry
Conserved sequence
Deoxyribonucleic acid
DNA
DNA - analysis
DNA - chemical synthesis
DNA biosynthesis
DNA probes
DNA sequencing
Emission
Fluorescence
Fragments
Nucleic Acid Amplification Techniques
Nucleic Acid Conformation
Nucleotide sequence
Nucleotides
Nucleotides - chemistry
Single-nucleotide polymorphism
Substrates
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Summary: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.
ISSN:0003-2700
1520-6882
DOI:10.1021/acs.analchem.0c00707