[Separation of DNA fragments from halobacterium halobium by capillary electrophoresis with non-gel sieving media].
The optimum electrophoresis separation conditions of Lambda DNA/EcoR I + Hind III fragments were investigated using the capillary column DB-1 coated with polysiloxane. When hydroxyethylcellulose (HEC, 1.0%, mass percentage) was used as the only non-gel sieving medium solution, some Lambda DNA fragments could not be separated. After polyvinylpyrrolidone (PVP, 2%, mass percentage) being added into the medium solution, PVP and HEC formed a network colloidal solution in buffer, and changed the network hole size of sieving media. In addition, the results showed that PVP could restrain the adsorption of capillary to DNA, reduced electroosmotic flow, and improved the selectivity of separation. For the first time, all fragments of Lambda DNA marker was separated completely under the same condition of mixed sieving medium solution. The method was applied to separate two groups of DNA fragments of halobacterium halobium, and the base pair number was conjectured.
Wang
Y C
YC
Chemistry Department, Wuhan University, Wuhan 430072, China. Wangyc@public.xn.hb.cn
Xiong
Y
Y
Zeng
Z R
ZR
Cheng
J K
JK
Shen
P
P
D002482:Cellulose / Q000031:analogs & derivatives*; D004269:DNA, Bacterial / Q000032:analysis*; D019075:Electrophoresis, Capillary / Q000295:instrumentation* / Q000379:methods; D019617:Halobacterium salinarum / Q000235:genetics*; D011205:Povidone; D012833:Siloxanes
D004269:DNA, Bacterial; D012833:Siloxanes; D002482:Cellulose; C002283:hydroxyethylcellulose; D011205:Povidone