Feulgen cytophotometry of pine nuclei. II. Effect of pectinase used in cell separation.
Pectinase used for cell separation prior to cytophotometry contains a DNase that is able to penetrate the cells of pine root tips and attack nuclear DNA. When pine root tips were exposed to 1% pectinase (pH 6.0), there was a decrease in nuclear DNA content at every sample point and a sharp drop between 16 and 20 hr. The effect of the DNase was eliminated by preparing the enzyme solution in 0.01 M sodium citrate or 0.001 M EDTA. It is suggested that heat denaturation of the DNase should also be effective and might be used in combination with the magnesium chelators.
D002469:Cell Separation / Q000379:methods; D004247:DNA / Q000032:analysis*; D003851:Deoxyribonucleases / Q000494:pharmacology; D006026:Glycoside Hydrolases / Q000494:pharmacology*; D010783:Photometry / Q000295:instrumentation / Q000379:methods; D011096:Polygalacturonase / Q000494:pharmacology*; D012639:Seeds / Q000032:analysis* / Q000166:cytology / Q000187:drug effects; D013194:Staining and Labeling / Q000379:methods
D004247:DNA; D003851:Deoxyribonucleases; D006026:Glycoside Hydrolases; D011096:Polygalacturonase