Analyses of Nuclear Proteins and Nucleic Acid Structures Using Atomic Force Microscopy.
Nuclear Bodies and Noncoding RNAs
Methods in molecular biology, January 2015
Jamie L Gilmore, Aiko Yoshida, Hirohide Takahashi, Katashi Deguchi, Toshiro Kobori, Emilie Louvet, Masahiro Kumeta, Shige H Yoshimura, Kunio Takeyasu, Jamie L. Gilmore, Shige H. Yoshimura
Since the inception of atomic force microscopy (AFM) in 1986, the value of this technology for exploring the structure and biophysical properties of a variety of biological samples has been increasingly recognized. AFM provides the opportunity to both image samples at nanometer resolution and also measure the forces on the surface of the sample. Here, we describe a variety of methods for studying nuclear samples including single nucleic acid molecules, higher-order chromatin structures, the nucleolus, and the nucleus. Protocols to prepare nucleic acids, nucleic acid-protein complexes, reconstituted chromatin, the cell nucleus, and the nucleolus are included, as well as protocols describing how to prepare the AFM substrate and the AFM tip. Finally, we describe how to perform conventional imaging, high-speed imaging, recognition imaging, force spectroscopy, and nanoindentation experiments.
|Members of the public||1||100%|
|Readers by professional status||Count||As %|
|Student > Bachelor||3||19%|
|Student > Ph. D. Student||3||19%|
|Student > Master||2||13%|
|Readers by discipline||Count||As %|
|Agricultural and Biological Sciences||7||44%|
|Biochemistry, Genetics and Molecular Biology||6||38%|
|Immunology and Microbiology||1||6%|
|Medicine and Dentistry||1||6%|