Direct visualisation of plasmid DNA in individual chromatography adsorbent particles by confocal scanning laser microscopy.
Confocal microscopy was used for the measurement of plasmid DNA adsorbed to individual adsorbent particles intended for anion-exchange and triple helix affinity chromatography. Plasmid DNA was visualized with the fluorescent dye YOYO-1, that forms a highly fluorescent complex with double stranded DNA. Confocal images were translated into fluorescence intensity profiles and the distribution of plasmid DNA in the particles was measured. The results that adsorption of plasmid DNA mainly takes place in an outer layer of the particles. The described procedure can also be advantageously used to demonstrate triple helix formation between plasmid DNA and immobilized oligonucleotides.
Ljunglöf
A
A
Amersham Pharmacia Biotech, Uppsala, Sweden. anders.ljunglof@eu.apbiotech.com
Bergvall
P
P
Bhikhabhai
R
R
Hjorth
R
R
D000327:Adsorption; D002845:Chromatography / Q000379:methods*; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D004247:DNA / Q000032:analysis*; D018613:Microscopy, Confocal; D010316:Particle Size; D010957:Plasmids / Q000235:genetics*
D004247:DNA