[Influence of sample dilution on separation and detection of DNA fragments by capillary electrophoresis].
Capillary electrophoresis has become an important and useful method to separate and determine DNA fragments. In molecular biochemistry, the volume of DNA sample is very small (microL level) and DNA sample is liable to be contaminated and degraded. According to theoretical inference and experiments, we propose that dilution of DNA sample solution can increase separation efficiency and resolution without evidently reducing height of peaks. By this method, the usage efficiency of DNA sample can be improved. It is also demonstrated the separation and detection of DNA fragments by capillary electrophoresis with hydroxyethyl cellulose non-gel sieving matrix and with laser-induced fluorescence charge-coupled device as detector. By using lower concentration non-gel matrix (0.4%), all 8 larger size fragments of lambda DNA/Hind III (125 bp-23 130 bp) can be completely separated. Twenty smaller size fragments of pBR322-Hae III DNA (18 bp-587 bp) can be separated by higher concentration (1.6%) non-gel matrix. As ratio of sample dilution is 10, two adjacent fragment (123 bp and 124 bp) of pBR322-Hae III DNA can be separated.
Song
L
L
Department of Chemistry, Wuhan University, Wuhan, 430072.
Chen
H
H
Zhang
L
L
Cheng
J
J
D004247:DNA / Q000032:analysis / Q000302:isolation & purification*; D019075:Electrophoresis, Capillary; D013048:Specimen Handling / Q000379:methods
D004247:DNA