Label-free monitoring of site-specific DNA cleavage by EcoRI endonuclease using cyclic voltammetry and electrochemical impedance.
Key Laboratory of Analytical Chemistry for Life Science of Shaanxi Province, School of Chemistry and Materials Science, Shaanxi Normal University, Xi'an 710062, PR China.
Analytica chimica acta 2009 Feb 16Cyclic voltammetry (CV) combined with electrochemical impedance spectroscopy (EIS) were proposed to monitor the site-specific DNA cleavage by EcoRI endonuclease without using external label. The alteration of CV and EIS signal demonstrated that double-strands (dsDNA) contain recognition sequence was cleaved by EcoRI endonuclease. Real-time monitoring indicated that the dsDNA was cleaved by EcoRI more than 90% after 2h of enzyme digestion time. Control experiment showed that the DNA cleavage by EcoRI endonuclease is site-specific for DNA sequence. Experimental results demonstrated that the efficiency of EcoRI cleavage was highly dependent on the concentration of EcoRI concentration in the range from 0.04 to 0.4UmuL(-1) with one almost linear relationship.
Citation
Yan Jin. Label-free monitoring of site-specific DNA cleavage by EcoRI endonuclease using cyclic voltammetry and electrochemical impedance. Analytica chimica acta. 2009 Feb 16;634(1):44-8
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PMID: 19154808
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