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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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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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creator	Sun, Feifei Zhao, Shengnan Peng, Manshu Fu, Qiang Gao, Huimin Jia, Yijing Na, Na Ouyang, Jin
description	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.
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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
title	Sequencing of Small DNA Fragments with Aggregated-Induced-Emission Molecule-Labeled Nucleotides
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