Visualization of DNA and RNA molecules, and protein-DNA complexes for electron microscopy.
This article provides step-by step instructions for the preparation of double- and single-stranded DNA and RNA molecules and protein-DNA complexes for electron microscopy (EM). Absorption, spreading, staining, dark-field imaging, and metal shadowing techniques are described in detail. A number of examples are illustrated on analysis of DNA replication, DNA repair and DNA recombination to demonstrate the usefulness of the technique for EM visualisation. Application of immunogold labeling of specific protein in DNA-protein complexes is also covered.
Hajibagheri
M A
MA
Electron Microscopy Unit, Imperial Cancer Research Fund, London, UK.
D000818:Animals; D022202:Chromosomes, Artificial, Bacterial / Q000648:ultrastructure; D004247:DNA / Q000648:ultrastructure*; D004260:DNA Repair; D004261:DNA Replication; D004268:DNA-Binding Proteins / Q000648:ultrastructure; D007150:Immunohistochemistry; D008854:Microscopy, Electron / Q000379:methods*; D010063:Ovum / Q000648:ultrastructure; D010957:Plasmids / Q000528:radiation effects / Q000648:ultrastructure; D011506:Proteins / Q000648:ultrastructure*; D012313:RNA / Q000648:ultrastructure*; D051135:Rad51 Recombinase; D051721:Rad52 DNA Repair and Recombination Protein; D011938:Rec A Recombinases / Q000648:ultrastructure; D011995:Recombination, Genetic; D014981:Xenopus / Q000502:physiology; D029867:Xenopus Proteins
D004268:DNA-Binding Proteins; D011506:Proteins; D051721:Rad52 DNA Repair and Recombination Protein; D029867:Xenopus Proteins; D012313:RNA; D004247:DNA; C495152:RAD51 protein, Xenopus; D051135:Rad51 Recombinase; D011938:Rec A Recombinases