Application of short monolithic columns for fast purification of plasmid DNA.
Anion-exchange chromatography is one of the most important methods in downstream processing of plasmid DNA, both as a process and as an analytical technique. Separation of plasmid DNA on traditional particle-based anion-exchange supports is usually slow. Moreover, such supports have a low capacity for plasmid DNA due to the steric exclusion effects. In this work, the separation of plasmid DNA using short monolithic columns, Convective Interaction Media, will be presented. It will be demonstrated that plasmid DNA can be purified from bacterial cells using alkaline lysis followed by chromatography on a very short weak anion-exchange chromatographic columns-disks-with good purity and quality within a short time. Furthermore, the separation of plasmid DNA from cell RNA can be carried out without the need of adding RNAse. Fast and efficient method for in-process control of the purified plasmid will be described as well.
Branovic
Karmen
K
Institute of Immunology Inc., Rockefeller Street 2, 10000 Zagreb, Croatia. kbranovic@imz.hr
Forcic
Dubravko
D
Ivancic
Jelena
J
Strancar
Ales
A
Barut
Milos
M
Kosutic Gulija
Tanja
T
Zgorelec
Renata
R
Mazuran
Renata
R
D000838:Anions; D002852:Chromatography, Ion Exchange / Q000295:instrumentation / Q000379:methods*; D004247:DNA / Q000302:isolation & purification*; D004587:Electrophoresis, Agar Gel; D004926:Escherichia coli / Q000235:genetics; D004996:Ethidium; D010957:Plasmids / Q000235:genetics*; D014169:Transformation, Bacterial
D000838:Anions; D004247:DNA; D004996:Ethidium