DNA orientation using specific avidin-ferritin biotin end labelling.
The orientation of DNA molecules has been determined by labelling one of the molecule end with a Biotin-labelled analog of dTTP (Bio-dUTP) and then by complexing the Bio-dUTP with Avidin-Ferritin. DNAs of phi X174, pBR322 and SV40 were end labelled with Bio-dUTP and imaged by Electron Microscopy (EM). This is a rapid, general method to unambiguously determine the orientation of DNA molecules for precise mapping and quantification of DNA secondary structures or protein-DNA interaction sites using EM.
D001360:Avidin / Q000031:analogs & derivatives; D010584:Bacteriophage phi X 174; D001710:Biotin / Q000031:analogs & derivatives; D004262:DNA Restriction Enzymes; D004269:DNA, Bacterial / Q000648:ultrastructure*; D004279:DNA, Viral / Q000648:ultrastructure*; D005293:Ferritins / Q000031:analogs & derivatives; D007202:Indicators and Reagents; D008854:Microscopy, Electron; D010957:Plasmids; D013539:Simian virus 40
D004269:DNA, Bacterial; D004279:DNA, Viral; D007202:Indicators and Reagents; C051805:avidin-ferritin biotin; D001360:Avidin; D001710:Biotin; D005293:Ferritins; D004262:DNA Restriction Enzymes