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Influence of Nucleotide-biased Fluorescence Emissions of SYBR Green II on the Result Consistence of Rolling Circle Amplification
Zhang Bin1; Jiang Jiquan1; Yuan Ying1; Guan Yifu1
2019
Source PublicationCHEMICAL RESEARCH IN CHINESE UNIVERSITIES
ISSN1005-9040
Volume35Issue:6Pages:1119-1123
AbstractThe fluorescence dye SYBR Green II(SG II) has been frequently used in rolling circle amplification(RCA) based analyses of nucleic acids. However, a good amount of inconsistencies have been reported in regards the quality and reproducibility of RCA reactions. To properly examine this experimental issue, here we utilized a series of synthetic oligonucleotides and circular templates to investigate the impact of SG II in RCA reactions. The results indicate that SG II enables a strong fluorescence signal only when complexing with guanosine(G) residue. In RCA reactions, long single-stranded RCA products, enriched with G residues, result in higher fluorescence emission when compared with the addition of other nucleotide residues. These results suggest that the nucleotide composition of the reaction can affect the amplification results and, eventually, can lead to inconsistent fluorescence of the RCA products. This work indicates that particular attention should be given when circular templates are designed for the quantitative analysis of nucleic acids, to further allow the signal reproducibility of RCA-based experiments.
KeywordDNA DETECTION QUADRUPLEX IDENTIFICATION STABILITY Rolling circle amplification SYBR Green II Nucleotide-biased Fluorescence emission
Indexed ByCSCD
Language英语
Funding Project[National Natural Science Foundation of China]
CSCD IDCSCD:6626732
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Document Type期刊论文
Identifierhttp://ir.imr.ac.cn/handle/321006/155543
Collection中国科学院金属研究所
Affiliation1.中国科学院地质与地球物理研究所
2.中国科学院金属研究所
Recommended Citation
GB/T 7714
Zhang Bin,Jiang Jiquan,Yuan Ying,et al. Influence of Nucleotide-biased Fluorescence Emissions of SYBR Green II on the Result Consistence of Rolling Circle Amplification[J]. CHEMICAL RESEARCH IN CHINESE UNIVERSITIES,2019,35(6):1119-1123.
APA Zhang Bin,Jiang Jiquan,Yuan Ying,&Guan Yifu.(2019).Influence of Nucleotide-biased Fluorescence Emissions of SYBR Green II on the Result Consistence of Rolling Circle Amplification.CHEMICAL RESEARCH IN CHINESE UNIVERSITIES,35(6),1119-1123.
MLA Zhang Bin,et al."Influence of Nucleotide-biased Fluorescence Emissions of SYBR Green II on the Result Consistence of Rolling Circle Amplification".CHEMICAL RESEARCH IN CHINESE UNIVERSITIES 35.6(2019):1119-1123.
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