By Jeffrey H. Miller
This quantity collates in a single resource method for in vivo genetic engineering and for genetic research in quite a lot of micro organism. not just is Escherichia coli good lined, yet so are different rising bacterial platforms
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Additional resources for Bacterial Genetic Systems
Phage titers achieved by this procedure are usually 3 × 109 to 2 × 10l° phage/ml. Lysogen Induction. Lysogens containing a P1 cm cl. 6). The culture is then diluted with an equal volume of L broth containing 10 mM CaCI2 that was preheated to 50°, and the culture is vigorously aerated at 42 ° for 15 min. The temperature of the water bath is then reduced to 39° and aeration continued until visible cell lysis is detected, about 50-60 min after temperature shift-up. The lysate is treated with chloroform and debris removed by centrifugation.
These colonies are used to inoculate overnight cultures which are then screened to ensure that they are lysogens, h ci857 lysogens are immune to h superinfection at 32 ° and die at 42 °. The lysogen we frequently use in preparing a transducing lysate is NS490 (= N205; hb2 red3 ci857 Sam7). In principle, any bred3 ci857 Sam7 lysogen could be used. 32 R. A. Weisberg and A. Landy, in "Lambda II" (R. Hendrix, J. Roberts, F. Stahl, and R. ), p. 211. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, 1983.
Ikeda and J. I. Tomizawa, J. Mol. Biol. 14, 85 (1965). C. Hanks, B. Newman, I. R. Oliver, and M. Masters, Mol. Gen. Genet. 214, 523 (1988). Ikeda and J. I. Tomizawa, J. Mol. Biol. 14, 110 (1965). L. Taylor and C. D. Trotter, Bacteriol. Rev. 31, 332 (1967). 5 x 10 -5 0 to 10 -2 10 -7 to 10 -5 which markers are transduced can vary by as much as 30-fold, ranging from 3 x 10 -4 to 10 -5 transductants per infected donor cell (Table I). These differences are due primarily to variation in the efficiency of incorporation of transducing DNA into the chromosome of the recipient cell.