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By T. Bergan (Ed.)

Proven nearly 30 years in the past, Methods in Microbiology is the main prestigious sequence dedicated to suggestions and technique within the box. Now absolutely remodeled, revitalized, with a brand new structure and multiplied scope, Methods in Microbiology will proceed to supply you with attempted and verified, state-of-the-art protocols to at once profit your learn.

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Mix and warm in a SO-60°C water bath. Lysis will be apparent by a clearing of the suspension and an increase in the viscosity. When the cells are SDS sensitive, treat all of the cells as described for the aliquots. 5 h before harvesting). (6) N-acetylmuramidase. This enzyme was isolated by Yokogawa et al. , (1975). 5. Add enzyme to a level of about S p g ml-' and incubate at 50°C. Again, remove aliquots and test for SDS sensitivity. ( c ) Lysostaphin. This enzyme appears to be specific for the rather unique cross-linking peptides in the cell walls of Staphylococcus (Schindler and Schuhardt, 1964).

25 ml of SDS. Swirl the flask, and warm in a 50-60°C water bath until lysis is complete. For organisms that are not disrupted by detergent alone, see Section A. 3 . Reduce the viscosity of the lysate by briefly subjecting it to sonic oscillation. Pronase b or proteinase K may be added at this time (50 p g ml-I). For some organisms, proteinase digestion is not needed, while for others it is essential, for example, where there is a high level of nuclease activity or, as in some cases, where the DNA appears to be closely associated with cell protein and will be removed with the denatured protein if not first released with proteinase.

12. Base composition of DNA as a function of the absorbance ratio of its acid dialysate. The broken line (a) represents the ratios of pure base-pair mixtures, and the solid line (b) represents the best line through 13 DNA samples whose mol% G + C values had been obtained from the literature or unpublished (these determinations were by chemical analysis, T , , or buoyant density). From Huang and Rosenberg (1966). 6. 0 While investigating the ultraviolet spectra of DNA and deoxyribonucleotides, Fredericq et al.

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