sybr-green-i and coralyne

Polydeoxyadenosine (poly (dA)) has been extensively applied for detecting many drug molecules. Herein, we developed a sensitive method for detecting coralyne and heparin using a modified DNA probe with poly (dA) at one end. In the absence of coralyne, the DNA probe was digested by the Exonuclease I (Exo I), and therefore the SYBR Green I (SG I) emitted an extremely low fluorescent signal. While coralyne specifically binding to poly (dA) with strong propensity could remarkably restrain the disintegration of the DNA probe, through which as a template the second strand of DNA sequence was formed with the introduction of DNA polymerase. Therefore, the fluorescent signal of SG I was intensified to quantify coralyne. Based on this method, heparin can be determined due to its strong affinity towards coralyne. This method showed a linear range from 2 to 500 nM for coralyne with a low detection limit of 0.98 nM, and the linear range of heparin was from 1 to 100 nM when 1.25 nm was the detection limit. The proposed method was also implemented successfully in biological samples and showed a potential application for screening potential therapeutic molecules.

Topics: Benzothiazoles; Berberine Alkaloids; Biosensing Techniques; Deoxyadenosines; Diamines; DNA; DNA Probes; Exodeoxyribonucleases; Heparin; Humans; Limit of Detection; Organic Chemicals; Quinolines

This study describes the development of a simple, sensitive, and selective-detection system for coralyne and polyadenylation reaction based on the fact that coralyne induces a conformational change of the polyadenosine [poly(A)] oligonucleotide through A-coralyne-A coordination, thereby enhancing the fluorescence of SYBR Green I.

Topics: Benzothiazoles; Berberine Alkaloids; Biosensing Techniques; Diamines; Fluorescent Dyes; Nucleic Acid Conformation; Oligonucleotides; Organic Chemicals; Polyadenylation; Quinolines; Repetitive Sequences, Nucleic Acid; Spectrometry, Fluorescence; Time Factors