The influence of polymer concentration, applied voltage, modulation depth and pulse frequency on DNA separation by pulsed field CE.
DNA fragments (0.1-10 kbp (kbp, kilo base pair)) separation by square-wave pulsed field CE in hydroxyethylcellulose (HEC, 1300 K) polymer was performed in this work. The effects of polymer concentration, pulse field strength, pulse frequency and modulation depth were investigated. We found that low HEC (about 0.1%) concentration is suitable for the separation of small DNA fragments (<1 kbp), whereas higher HEC concentration (>0.5%) is appropriated for high-mass DNA molecular (>1 kbp) separation. The mobility of DNA fragments is nearly linearly related to average separation voltage under pulsed field conditions. Higher modulation depth is suited to separate the longer DNA fragments and lower modulation depth favors the resolution of short DNA fragments. Thus, the intermediate modulation depth (100%) and pulse frequency (about 31.3 Hz) are prerequisite for high-resolution DNA separation.
Li
Zhenqing
Z
Department of Physics, Optical Engineering, Shanghai Jiao Tong University, Shanghai, P. R. China.
Dou
Xiaoming
X
Ni
Yi
Y
Sumitomo
Keiko
K
Yamaguchi
Yoshinori
Y
D002482:Cellulose / Q000031:analogs & derivatives / Q000737:chemistry; D004247:DNA / Q000737:chemistry / Q000302:isolation & purification*; D004560:Electricity; D055664:Electrochemical Techniques / Q000295:instrumentation / Q000379:methods; D019075:Electrophoresis, Capillary / Q000295:instrumentation / Q000379:methods; D011108:Polymers / Q000737:chemistry*
D011108:Polymers; D002482:Cellulose; C002283:hydroxyethylcellulose; D004247:DNA