DNA aggregation and cleavage in CGE induced by high electric field in aqueous solution accompanying electrokinetic sample injection.
The phenomenon of peak area decrease due to high injection voltage (Vinj , e.g. 10-30 kV, 200-600 V/cm in the 50 cm capillary) was found in the analysis of very dilute DNA fragments (<0.2 mg/L) by using high-sensitive electrokinetic supercharging-CGE. The possibility of DNA cleavage in aqueous solution was suggested, in addition to the aggregation phenomenon that is already known. The analysis of intentionally voltage-affected fragments (at 200 V/cm) also showed decreased peak areas depending on the time of the voltage being applied. Computer simulation suggested that a high electric field (a few kV/cm or more) could be generated partly between the electrode and the capillary end during electrokinetic injection (EKI) process. After thorough experimental verification, it was found that the factors affecting the damage during EKI were the magnitude of electric field, the distance between tips of electrode and capillary (De/c ), sample concentration and traveling time during EKI in sample vials. Furthermore, these factors are correlating with each other. A low conductivity of diluted sample would cause a high electric field (over a few hundred volts per centimeter), while the longer De/c results in a longer traveling time during EKI, which may cause a larger degree of damage (aggregation and cleavage) on the DNA fragments. As an important practical implication of this study, when the dilute DNA fragments (sub mg/L) are to be analyzed by CGE using EKI, injection voltage should be kept as low as possible.
Applied Chemistry, Department of Chemistry and Chemical Engineering, Graduate School of Engineering, Hiroshima University, Japan.
Xiaoxue
X
Ye
Satomi
S
Mori
Zhongqi
Z
Xu
Shinjiro
S
Hayakawa
Takeshi
T
Hirokawa
D020374:Chromatography, Micellar Electrokinetic Capillary / Q000379:methods*; D004247:DNA / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification / Q000528:radiation effects*; D004249:DNA Damage / Q000528:radiation effects; D004553:Electric Conductivity; D004560:Electricity; D019075:Electrophoresis, Capillary / Q000379:methods*
D004247:DNA
CE; DNA analysis; DNA damage; Electrokinetic injection; Electrokinetic supercharging; Injection voltage