Label-free monitoring of site-specific DNA cleavage by EcoRI endonuclease using cyclic voltammetry and electrochemical impedance.
Cyclic 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.
Jin
Yan
Y
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.
D001665:Binding Sites; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D053837:DNA Cleavage; D015246:Deoxyribonuclease EcoRI / Q000737:chemistry / Q000378:metabolism*; D004563:Electrochemistry / Q000379:methods*; D013379:Substrate Specificity; D013997:Time Factors
D015246:Deoxyribonuclease EcoRI