Chromatographic behaviour and purification of linear  lambda
phage  and plasmid DNA molecules on 2-hydroxyethyl
methacrylate-ethylene dimethacrylate-based supports

J 2003

Chromatographic behaviour and purification of linear lambda phage and plasmid DNA molecules on 2-hydroxyethyl methacrylate-ethylene dimethacrylate-based supports

RITTICH, Bohuslav, Alena ŠPANOVÁ, Magdalena SKALNÍKOVÁ and Milan BENEŠ

Basic information

Original name

Chromatographic behaviour and purification of linear lambda phage and plasmid DNA molecules on 2-hydroxyethyl methacrylate-ethylene dimethacrylate-based supports

Authors

RITTICH, Bohuslav (203 Czech Republic, guarantor), Alena ŠPANOVÁ (203 Czech Republic), Magdalena SKALNÍKOVÁ (203 Czech Republic) and Milan BENEŠ (203 Czech Republic)

Edition

Journal of Chromatography A, Amsterdam, Elsevier, 2003, 0021-9673

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10406 Analytical chemistry

Country of publisher

Netherlands

Confidentiality degree

není předmětem státního či obchodního tajemství

Impact factor

Impact factor: 2.922

RIV identification code

RIV/00216224:14310/03:00008144

Organization unit

Faculty of Science

Keywords in English

Stationary phases; LC; slalom chromatography; DNA; hydroxyethyl methacrylate-ethylene dimethacrylate

Abstract

V originále

The HEMA-BIO 1000 supports based on a copolymer of 2-hydroxyethyl methacrylate and ethylene dimethacrylate were used for separation of lambda DNA and its fragments and plasmid pBR322 DNA. The separation of fragments greater than 6.6 kbp was demonstrated according to the slalom chromatography mechanism on column for gel permeation chromatography in the case of linear lambda DNA fragments. The influence of the particle size of column packing, mobile phase rate, and KCl concentration in mobile phase was discussed. The purification of plasmid DNA pBR322 using GPC was more rapid in comparison with gel electrophoresis. The presence of salts in the eluate is not disadvantageous. DNA can be recovered from the eluate by ethanol precipitation. Plasmid DNA pBR322 isolated in this way was suitable for different biological applications (cleavage with restrictases, electrotransformation into bacterial cells).

Links

GA203/98/1231, research and development project

Name: Orientovaná imobilizace enzymů a protilátek s využitím biospecifických interakcí

Investor: Czech Science Foundation, Oriented immobilization of enzymes and antibodies using biospecific interactions

Citovat

RITTICH, Bohuslav, Alena ŠPANOVÁ, Magdalena SKALNÍKOVÁ and Milan BENEŠ. Chromatographic behaviour and purification of linear lambda phage and plasmid DNA molecules on 2-hydroxyethyl methacrylate-ethylene dimethacrylate-based supports. Journal of Chromatography A. Amsterdam: Elsevier, 2003, vol. 1009, 1-2, p. 207-214. ISSN 0021-9673.

@article{488218,
   author = {Rittich, Bohuslav and Španová, Alena and Skalníková, Magdalena and Beneš, Milan},
   article_location = {Amsterdam},
   article_number = {1-2},
   keywords = {Stationary phases; LC; slalom chromatography; DNA; hydroxyethyl methacrylate-ethylene dimethacrylate},
   language = {eng},
   issn = {0021-9673},
   journal = {Journal of Chromatography A},
   title = {Chromatographic behaviour and purification of linear lambda phage and plasmid DNA molecules on 2-hydroxyethyl methacrylate-ethylene dimethacrylate-based supports},
   volume = {1009},
   year = {2003}
}

TY  - JOUR
ID  - 488218
AU  - Rittich, Bohuslav - Španová, Alena - Skalníková, Magdalena - Beneš, Milan
PY  - 2003
TI  - Chromatographic behaviour and purification of linear lambda phage and plasmid DNA molecules on 2-hydroxyethyl methacrylate-ethylene dimethacrylate-based supports
JF  - Journal of Chromatography A
VL  - 1009
IS  - 1-2
SP  - 207-214
EP  - 207-214
PB  - Elsevier
SN  - 00219673
KW  - Stationary phases
KW  - LC
KW  - slalom chromatography
KW  - DNA
KW  - hydroxyethyl methacrylate-ethylene dimethacrylate
N2  - The HEMA-BIO 1000 supports based on a copolymer of 2-hydroxyethyl methacrylate and ethylene dimethacrylate were used for separation of lambda DNA and its fragments and plasmid pBR322 DNA. The separation of fragments greater than 6.6 kbp was demonstrated according to the slalom chromatography mechanism on column for gel permeation chromatography in the case of linear lambda DNA fragments. The influence of the particle size of column packing, mobile phase rate, and KCl concentration in mobile phase was discussed. The purification of plasmid DNA pBR322 using GPC was more rapid in comparison with gel electrophoresis. The presence of salts in the eluate is not disadvantageous. DNA can be recovered from the eluate by ethanol precipitation. Plasmid DNA pBR322 isolated in this way was suitable for different biological applications (cleavage with restrictases, electrotransformation into bacterial cells).
ER  -

RITTICH, Bohuslav, Alena ŠPANOVÁ, Magdalena SKALNÍKOVÁ and Milan BENEŠ. Chromatographic behaviour and purification of linear lambda phage and plasmid DNA molecules on 2-hydroxyethyl methacrylate-ethylene dimethacrylate-based supports. \textit{Journal of Chromatography A}. Amsterdam: Elsevier, 2003, vol.~1009, 1-2, p.~207-214. ISSN~0021-9673.

Detailed Information on Publication Record