TY  - JOUR
AU  - Frerix, A.
AU  - Geilenkirchen, P.
AU  - Müller, M.
AU  - Kula, M.-R.
AU  - Hubbuch, J.
TI  - Separation of Genomic DNA, RNA, and Open Circular Plasmid DNA From Supercoiled Plasmid DNA by Combining Denaturatin, Selective Renaturation and Aqueous Two-Phase Extraction
JO  - Biotechnology & bioengineering
VL  - 96
SN  - 0006-3592
CY  - New York, NY [u.a.]
PB  - Wiley
M1  - PreJuSER-56151
SP  - 57 - 66
PY  - 2007
N1  - Record converted from VDB: 12.11.2012
AB  - In the current study we developed a process for the capture of pDNA exploiting the ability of aqueous two-phase systems to differentiate between different forms of DNA. In these systems scpDNA exhibits a near quantitative partitioning in the salt-rich bottom phase. The successive recovery from the salt rich bottom phase is accomplished by a novel membrane step. The polish operation to meet final purity demands is again based on a system exploiting a combination of the denaturation of the nucleic acids present, specific renaturation of scpDNA, and an ATP system able to differentiate between the renatured scpDNA and the denatured contaminants such as ocpDNA and genomic host DNA. This polish step thus allows a rapid and efficient separation of scpDNA from contaminating nucleic acids which up to date otherwise only can be accomplished with much more cumbersome chromatographic methods. In a benchmark comparison, it could be shown that the newly developed process exhibits a comparable yield to an industrial standard process while at the same time showing superior performance in terms of purity and process time. Additionally it could be shown that the developed polish procedure can be applied as a standalone module to support already existing processes.
KW  - Chemical Fractionation: methods
KW  - DNA: genetics
KW  - DNA: isolation & purification
KW  - DNA, Circular: genetics
KW  - DNA, Circular: isolation & purification
KW  - Electrophoresis, Agar Gel: methods
KW  - Phase Transition
KW  - Plasmids: genetics
KW  - Plasmids: isolation & purification
KW  - RNA: genetics
KW  - RNA: isolation & purification
KW  - Ultrafiltration: methods
KW  - Water: chemistry
KW  - DNA, Circular (NLM Chemicals)
KW  - RNA (NLM Chemicals)
KW  - Water (NLM Chemicals)
KW  - DNA (NLM Chemicals)
KW  - J (WoSType)
LB  - PUB:(DE-HGF)16
C6  - pmid:16937402
UR  - <Go to ISI:>//WOS:000243176500007
DO  - DOI:10.1002/bit.21166
UR  - https://juser.fz-juelich.de/record/56151
ER  -