By Carl Wu, C. David Allis
DNA within the nucleus of plant and animal cells is kept within the kind of chromatin. Chromatin and the Chromatin remodelling enzymes play a massive position in gene transcription.
Read Online or Download Methods in Enzymology, Vol. 376: Chromatin and Chromatin Remodeling Enzymes, Part B PDF
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Additional resources for Methods in Enzymology, Vol. 376: Chromatin and Chromatin Remodeling Enzymes, Part B
The ultimate goal is to understand the different states of chromatin at an atomic level of resolution, but the range of chromatin structures accessible to X-ray crystallography is very limited. ’’ The range of transmission EM-based technologies available for visualizing chromatin and chromatin remodeling complexes is listed in Table I. For each method, we provide experimental guidelines based on recent work in our laboratory. General Considerations With any imaging technique, the information retrievable is limited by the quality of the sample, and for chromatin, the most important considerations are as follows.
Edu). . com). Electron Microscope Data Collection CCD Cameras Versus Film The theoretical pros and cons of digital versus film recording of data have been extensively discussed8; here we will consider some of the practical considerations. For beam-sensitive specimens (negatively stained or frozen hydrated) the microscope must be able to record low-dose images via deflection/shuttering of the beam above the sample. For most samples 6 J. Bednar and C. L. Woodcock, Meth. Enz. 304, 191 (1999). C.
14 If the number of images comprising each class is very small, indicating either sample or staining heterogeneity, the half-averages will be very noisy, and the resolution may be underestimated. Accuracy of the Reconstruction The final question becomes how to determine if the reconstruction is correct. For a wholly asymmetric object, the reconstruction algorithms might be producing only one of many possible solutions and there is no quantitative way to determine if the one generated is the correct one.