Applications of Chimeric Genes and Hybrid Proteins by Thorner J. (ed.), Emr S.D. (ed.), Abelson J.N. (ed.)

By Thorner J. (ed.), Emr S.D. (ed.), Abelson J.N. (ed.)

The significantly acclaimed laboratory common for greater than 40 years, equipment in Enzymology is among the such a lot hugely revered guides within the box of biochemistry. because 1955, every one quantity has been eagerly awaited, usually consulted, and praised via researchers and reviewers alike. Now with greater than three hundred volumes (all of them nonetheless in print), the sequence comprises a lot fabric nonetheless appropriate today-truly a necessary booklet for researchers in all fields of existence sciences.

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LI Typically, the laser power does not need to exceed 100 mW (higher laser power contributes to higher noise levels in the forward scatter detectors). FACS are more specialized machines that are often available as shared equipment in immunology core facilities and are often operated by a full-time trained technician, who may not be accustomed to performing bacterial cell sorts. It is recommended that the researcher discuss the experimental setup and objectives with the FACS operator prior to experimentation.

This rich medium, which was originally formulated to facilitate screening for Lac" gram-negative microorganisms, contains bile salts (to inhibit the growth of nonenteric bacteria) and neutral red as a pH indicator. Lac bacteria, which can grow on lactose MacConkey agar, form white colonies. In contrast, cells that can ferment lactose form acid by-products, which turn the pH-sensitive dye red, and thus produce red colonies. Unlike X-Gal, phenol red is a diffusible dye, and therefore the colony phenotype should be read promptly after incubation, as differences will become less clear over time.

R. Parks, L. A. Herzenberg, and L. A. Herzenberg, in "Fundamental Immunology" (W. E. ), p. 781. Raven Press, New York, 1989. 9 R. H. Valdivia and S. Falkow, MoL Microbiol. 22, 367 (1996). 50 GENE VUSIONS [41 Bacteria bearing a GFP promoter trap library stimulus -Low pH -Macrophages -Host tissues . + o o o c o Collect fluorescent bacteria No Collect fluorescent . --, bacteria Stnmulus- I activated[. ,- ~ stimulus IT I o o Collect + o,-. n0n-flu0rescent bacteria F~G. 1. Fluorescence-based selection strategy for the identification of differentially expressed genes.

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