Rapid method for purification of plasmid DNA and DNA fragments from DNA linkers using high-performance liquid chromatography on TSK-PW gel
1982; Elsevier BV; Volume: 240; Issue: 1 Linguagem: Inglês
10.1016/s0021-9673(01)84016-4
ISSN1873-3778
AutoresMichael E. Himmel, Peter J. Perna, Michael McDonell,
Tópico(s)Bacteriophages and microbial interactions
ResumoHigh-performance size exclusion chromatography (HPSEC) using TSK-G5000 PW in Tri buffer has been found to be a reliable method for the rapid fractionation of DNA ligation products. Plasmid and fragmented phage DNAs were found to elute in less than 2 min with recoveries greater than 98%. Escherichia coli transfection studies, using plasmid DNA that had been subjected to HPSEC column fractionation, showed high transformation efficiencies. MgCl2, a component of the DNA ligation reaction, was found to produce DNA-column support interactions, which resulted in low DNA recoveries. Such interactions were eliminated by chelation with ethylenediaminetetraacetate prior to chromatography.
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