Premium
The influence of polymer concentration, applied voltage, modulation depth and pulse frequency on DNA separation by pulsed field CE
Author(s) -
Li Zhenqing,
Dou Xiaoming,
Ni Yi,
Sumitomo Keiko,
Yamaguchi Yoshinori
Publication year - 2010
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201000188
Subject(s) - pulse (music) , modulation (music) , square wave , resolution (logic) , dna , analytical chemistry (journal) , chemistry , pulse width modulation , materials science , voltage , chromatography , physics , biochemistry , quantum mechanics , artificial intelligence , computer science , acoustics
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.