Studies on the retention of plasmid DNA and Escherichia coli nucleic acids by hydrophobic interaction chromatography.
This work presents studies on the interactions of supercoiled plasmid DNA and Escherichia coli genomic DNA (gDNA) and RNA, with an hydrophobic interaction chromatography (HIC) gel, obtained by derivatisation of Sepharose CL-6B with 1,4-butanediol diglycidyl ether. Nucleic acids purified from E. coli were injected separately in the above HIC column and eluted with 1.5 M (NH4)2SO4 in the buffer. The column was able to separate single-stranded from double-stranded nucleic acids. RNA and denatured gDNA were retarded in a different way due to the interactions of the exposed hydrophobic bases with the ligands. Supercoiled plasmid DNA, on the contrary, eluted in the flowthrough.
Diogo
M M
MM
Centro de Engenharia Biológica e Química, Instituto Superior Técnico, Lisboa, Portugal.
Queiroz
J A
JA
Prazeres
D M
DM
D002072:Butylene Glycols; D002847:Chromatography, Agarose / Q000379:methods*; D004269:DNA, Bacterial / Q000302:isolation & purification; D004278:DNA, Superhelical / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000737:chemistry*; D010957:Plasmids / Q000302:isolation & purification*; D012329:RNA, Bacterial / Q000302:isolation & purification; D012685:Sepharose / Q000031:analogs & derivatives*
D002072:Butylene Glycols; D004269:DNA, Bacterial; D004278:DNA, Superhelical; D012329:RNA, Bacterial; C047264:sepharose CL 6B; D012685:Sepharose; C014376:1,4-bis(2,3-epoxypropoxy)butane