Read e-book online DNA'Protein Interactions: Principles and Protocols (Methods PDF

By Tom Moss

ISBN-10: 0896036251

ISBN-13: 9780896036253

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Extra resources for DNA'Protein Interactions: Principles and Protocols (Methods in Molecular Biology)

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Spin the tube for a few seconds in a microcentrifuge and transfer the liquid to a new tube. Avoid transferring gel pieces to the new tube. Add 200 µL of watersaturated neutralized phenol, shake 1 min, and centrifuge for 5 min at room temperature. Remove aqueous layer (upper), transfer it to a fresh Eppendorf tube, and add 30 µL of 3 M sodium acetate and 1 mL of ice-cold 100% ethanol. Shake the tube for a few seconds and put into –70°C for at least 30 min. 58 Zaychikov et al. 5. Spin the sample in a microcentrifuge for 30 min and remove the supernatant.

2. , step 2. 3. , step 5). 4. , steps 6 and 7. The half-life of the complex is in most cases not changed by the Exo III digestion of the DNA. 5. Expose the gel at –70°C to X-ray film using an intensifying screen. A 1-h exposure should be enough to recognize the complexed bands. If not, the recovered DNA will be insufficient for the subsequent sequencing gel analysis. 6. Before removing the film for development mark exactly its position on the gel. 7. Excise the complex bands of interest with a spatula.

Biol. 10, 1–9. Wilkison, W. , Min, H. , Claffey, K. , Satterberg, B. , and Spiegelman, B. M. (1990) Control of the adipisin gene in adipocyte differentiation. J. Biol. Chem. 265, 477–482. Further Reading Kow, Y. W. (1989) Mechanism of action of Escherichia coli Exonuclease III. Biochemistry 28, 3280–3287. Hydroxyl Radical Footprinting 49 5 Hydroxyl Radical Footprinting Evgeny Zaychikov, Peter Schickor, Ludmilla Denissova, and Hermann Heumann 1. Introduction The basic principle of the DNA footprinting technique is the measurement of accessibility of the DNA using a probe.

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DNA'Protein Interactions: Principles and Protocols (Methods in Molecular Biology) by Tom Moss


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