Elimination of non-specific nucleases from restriction endonuclease preparations by different binding on free DNA ligand.
In the purification of a novel restriction endonuclease (an AvaIII isoschizomer, isolated in this laboratory) standard methods were insufficient to eliminate non-specific nuclease contaminations. Taking advantage of the specific site recognition and binding of the restriction endonuclease on DNAs, a method is described for the simple extraction of non-specific nucleases. DNA substrates without recognizable sites do not bind the restriction endonuclease, while non-specific nucleases are absorbed to, and eliminated with, the DNA via gel filtration chromatography under special conditions.
Geck
P
P
Institute of Microbiology, Semmelweis University Medical School, Budapest, Hungary.
Molnár
A
A
Nász
I
I
D001667:Binding, Competitive; D002850:Chromatography, Gel; D004262:DNA Restriction Enzymes / Q000302:isolation & purification* / Q000378:metabolism; D004269:DNA, Bacterial / Q000378:metabolism*; D003851:Deoxyribonucleases / Q000378:metabolism*; D004587:Electrophoresis, Agar Gel; D004926:Escherichia coli / Q000235:genetics; D008024:Ligands
D004269:DNA, Bacterial; D008024:Ligands; D003851:Deoxyribonucleases; D004262:DNA Restriction Enzymes