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Three straight-sided Dewars, one for the cold trap described above and two for freezing cells. The size of the Dewars should be cylmdncal 15 cm diameter and 20 cm in length, inside dimensions Cooling apparatus. The tubes of cells are chilled at -20 to -30°C during drying (Note 1). This can be done using a refrigerated methanol bath such as the “Multicool” (FTS Systems, Stone Ridge, NY) Lacking that, the chillmg may be done m a freezer or the freezmg compartment of a refrigerator The door gasket of the freezer is cut to allow passage of the vacuum line.

0X SSC (seeNote 4) 18. X-ray film developer and fixer for the autoradlography are commercially avallable, and are made up according to the manufacturer’s mstructlons. 0), IM MgC12, and 10% (w/v) SDS should be prepared and can be stored at room temperature. Stock solutions of restriction enzyme buffers, bovme serum albumin, polyadenyllc acid, 100 x Denhardt’s, and herring sperm DNA should be stored at -20°C. Hybrldlzatlon solutions are prepared fresh as required. Method Restriction Encionuclease Digestion (See also Chapter 31) 1.

Matthews University of California, Department of Biological Chemistry, School of Medicine, Davis, California Introduction Autoradiography of gels containing a 32P label is used to detect and quantitate the radioactive label in a particular band or spot. Since 32P is a high energy p-emitting isotope, no fluor is required in the gel (as in fluorography) to increase the efficiency of detection. , calcium tungstate), where the radiation from the isotope is absorbed by a fluorescent compound that reemits the energy as light.

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Nucleic Acids by Walker J.M. (ed.)

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