Preparation of Culture Medium

It is a method of elemental analysis for various practical reasons and it is essentially suitable for analysis only of metals. A large number of elements can be analyzed for at trace levels. Therefore, its biotechnologic applications mainly involve in measurement of inorganic elements such as alkali, trace, and heavy metals in biological investigations. It is also used in industry to monitor contaminating inorganic elements in bioreactors fermentation process and preparation of culture media. 

Use Biochemical research, preparation of culture media, medicine. 

Use Biological and clinical studies, preparation of culture media, organic intermediate, ingredient of aspartame, detergents, fungicides, germicides, metal complexation. Available commercially as dl(—)-, l(+)-, and DL-aspartic acid. 

Preparation of Culture Media for Fertilized One-Celled Eggs... 

Table 2. Vitamin soiution used for the preparation of culture media for soil and water bacteria (after Schlegel). 2-3 ml of this solution are added to 1,000 ml of culture medium.

A. PREPARATION OF CULTURE MEDIA AND RELATED REAGENTS See Gordon and Ruddle (1983) and Quinn et al. (1982). 

A series of culture media was prepared, containing known amounts of gib-berellic acid, and assayed by the two methods described (Table III). 

To cultivate microorganisms, culture medium has to be prepared in one of the commonly employed culture vessels a test tube, a flask, a Petri dish, or a fermenter. There are two main types of culture media natural (or empirical, or complex) and synthetic (or chemically defined) media. They vary widely in form and composition, depending on the species of organism to be cultivated and the purpose of the cultivation. 

Ail reagents and culture media are recorded upon receipt or preparation. Reagents made up in the laboratory are prepared according to written procedures and appropriately labelled. Both positive and negative controls are applied to verify the suitability of culture media The size of the inoculum used in positive controls is appropriate to the sensitivity required. Records are maintained. 

Preparation of culture solutions and culture media General... 

Chicken embryo fibroblast cells (CEF) were prepared from the carcasses of 10-day-old embryos and maintained in complete growth medium as described [5]. The cells were used between the third and seventh subcultivation after initiation of the cultures. Cells were labeled in vivo with 125 juCi of [2,8,5 - H]adenosine per ml of culture media (50.5 Ci mmol 1 Ci = 3.7 x 10 Bq New England Nuclear) as described [5]. 

Standard methods for culture of anaerobic bacteria and preparation of anoxic media were u.sed throughout. Media and buffers were made anoxic by boiling and cooling under a stream of O -free N2 or N2 C()2 (80 20) gas. All reactions were performed in an anaerobic chamber (( oy I.aboratorics. Inc., (Irass Fake. Ml) with an... 

For a detailed discussion of culture media, see Cherbas and Cherbas (1998). The situation for casual cell users has been simplified by the fact that most common media for Drosophila cell culture are now commercially available. For example, most lines can be grown in M3 medium supplemented with 10% fetal calf serum, and M3 is available commercially both as liquid and as dry powder. For our larger-scale needs, we continue to manufacture M3 medium using the recipe in Schneider and Blumenthal (1978). Antibiotics are not in general necessary. For serum and its preparation by heat treatment and for more information about needles, see Cherbas and Cherbas (1998). 

A large number of different food media have been used for the culture of D. melanogaster. Several are described by Ashburner and Thompson (1978), Ashburner (1989b), Roberts and Standen (1998), and on the Bloomington Stock Center Web Site http //fly.bio.indi-ana.edu/media.recipes.htm. Only two are described below a general-purpose medium, and one suitable for growing larvae for polytene chromosome preparations. Table 35.2 lists the sources of ingredients used in the preparation of these media. In addition, instant media are available commercially (see Table 35.2). 

Fig. 5. Fermentative production of amino acids (140). A, pure culture B, inoculation C, boiler D, air compressor E, air filter F, seed tank G, ammonia water for pH control H, fermenter I, sterilizer , culture media K, preparation tank L, centrifugal separator M, ion-exchange column N, crystallizing...

The commercial production of mitomycin involves the preparation of mitomycin-containing broths by culturing a mitomycin-producing organism, e.g. Streptomyces caespitosus, in suitable media as described at length In the literature. At the end of the fermentation cycle the whole broth is usually centrifuged, filtered or otherwise treated to separate the solids (mycelia) from the supernatant which contains substantially all of the antibiotic activity. 

Figure 3.7 shows the growth of R. rubrum in a batch fermentation process using a gaseous carbon source (CO). The data shown follow the logistic model as fitted by (3.14.2.11) with the solid lines, which also represent an unstructured rate model without any lag phase. The software Sigma Plot was used to fit model (3.14.2.11) to the experimental data. An increase in concentration of acetate in the prepared culture media did not improve the cell dry weight at values of 2.5 and 3 gT-1 acetate, as shown in Figure 3.7. However, the exponential growth rates were clearly observed with acetate concentrations of 0.5-2 g-F1 hi the culture media.

A seed culture of S. cerevisiae ATCC 24860 (American Type Culture Collection, Manassas, VA, USA) was grown in a media of 5g glucose, and 0.5 g yeast extract, respectively, 1.5 g KH2P04 and 2.25 g Na2P04 phosphate buffer up to a total volume of distilled water, 500 ml. The media was sterilised at 121 °C for 15 min. The stock culture of the microorganisms was transferred to the broth media for preparation of seed culture. 

Seed culture of Saccharomyces cerevisiae (ATCC 24860) is grown in a rich medium comprising of 1 g glucose, 0.1 g peptone and yeast extract, 0.33 g KH2P04 and 0.03 g Na2HP04 in 100 ml distilled water. The media will be autoclaved at 126 °C and 15 psig for 20 min. The stock culture from ATCC media is transferred to a prepared seed culture. The pH of... 

To prepare CO solution for the experimental purpose, it is recommended to bubble 20 ml of stock solution in a sealed glass tube with a stream of pure CO gas. The bubbling process lasts for 20 min under the pressure of 100 kPa at 37°C [3]. One microliter of this CO-saturated solution is estimated to contain 30 ng of the gas based on the solubility of CO at 37°C, the extent of dilution of the CO-saturated solution, and the assumption that the loss of the added CO from the bath solution at the time of experiments is negligible. The stock solution of CO should be freshly prepared before each experiment and then should be diluted immediately to the desired concentration with the bath solution or culture media. 

For the evaluation of preparations to be used against pathogenic fungi, suitable cultures of these pathogens should be used. To test substances intended to inhibit general contaminants, cultures of common fungi obtained conveniently by exposing Petri dishes of solid media to the atmosphere may be used, or alternatively dust or soil may be used as a source of a mixed inoculum. 

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