Carl Wu, C. David Allis's Methods in Enzymology, Vol. 376: Chromatin and Chromatin PDF

By Carl Wu, C. David Allis

DNA within the nucleus of plant and animal cells is saved within the kind of chromatin. Chromatin and the Chromatin remodelling enzymes play a major position in gene transcription.

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Carl Wu, C. David Allis's Methods in Enzymology, Vol. 376: Chromatin and Chromatin PDF

DNA within the nucleus of plant and animal cells is kept within the kind of chromatin. Chromatin and the Chromatin remodelling enzymes play a big function in gene transcription.

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Additional info for Methods in Enzymology, Vol. 376: Chromatin and Chromatin Remodeling Enzymes, Part B

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Com). com). tvips. 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).

2. Wick off with filter paper. 3. Wash with 5 drops of ddH20, leaving the last drop on the grid for 30 s before wicking and air drying. 4. 3 l of colloidal gold solution (bead size 5–10 nm) is applied to the same side, wicked, and rinsed with 5 drops ddH2O and air-dried. 5. The sample is applied to the front surface of the grid and negatively stained as described above. The concentration of gold solution will need to be experimentally determined so that least 5 beads are present in the recorded image field.

It is often highly informative to view this z-series as a movie, to study transitions between structures in the interior of the volume. The volume can be sliced to 1-pixel thickness, to generate highly detailed views of substructure not visible in projections of the whole reconstructed volume. The most difficult aspect of tomography is understanding the 3D structure in the volumes. For chromatin, the use of isosurface contouring has proven unsatisfactory. The most useful information is accessed by viewing the entire volume as its original greyscale data, rendered with a transparency gradient to allow seeing into the volume.

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Methods in Enzymology, Vol. 376: Chromatin and Chromatin Remodeling Enzymes, Part B by Carl Wu, C. David Allis


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