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Extra resources for Metabolism of Amino Acids and Amines Part A
89, 915 (1965).  EXPERIMENTAL METHODS FOR Bacillus subtilis 45 C. Relative Effectiveness o f Mutagenic Treatments It is difficult at present to compile a list of mutagen effectiveness in vivo since each investigator has his own means for calculating mutant frequency. The most popular technique in the past was UV-irradiation (see Table II); however, NG treatment is gaining favor both because of its effectiveness and ease of manipulation. For in vitro mutagenesis, Bresler et al? 2. IV. Genetic Techniques A.
Nitrous Acid (HN02). Although used primarily in vitro, nitrous acid can be used in vivo to p r o d u c e mutations. T o a saturated culture in minimal m e d i u m , NaNO2 is a d d e d to give a 1 M concentration; the mixture is incubated at 25 ° for 1 hour, and the survivors are screened for mutants. 36 I n the in vitro system, mutagenesis is as effective with native DNA as it is with d e n a t u r e d DNA. 2 and treating for 30-60 minutes at 30-37 °. 5. T h e DNA is then used for transformation.
Analogs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36 38 39 42 43 45 45 45 49 49 50 51 51 53 53 54 54 55 55 56 56 57 57 59 I. Introduction Bacillus subtilis is actively being used for both biochemical and genetic experiments because of its prototrophic nature and the availability of genetic systems (transformation and transduction). It also has a spore stage which provides a simple model system for studying the problems of cell differentiation.