By J.R. Norris and D.W. Ribbons (Eds.)
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Additional resources for Methods in Microbiology, Vol. 3, Part B
25 g 3-5 g 10 g (c) The medium of Starkey (1935). 01 g (d) Neutral medium for nonaciduric species of Thiobacillus (Skerman, 1967). Use the Pope and Skerman Basic Mineral Salts Medium. Follow the preparation of the basal medium, except at Step 2 add 110 ml of water instead of 210, and at Step 7 make the final volume to 900ml instead of 1000. Sterilize at 121°C for 20 min. Cool. Then add aseptically 100 ml of sterile NazSz03 solution (Solution 5, Group A). The medium should be dispensed in a depth of not more than 1 cm in Erlenmeyer flasks.
Prepare the silicic acid by the method of Pramer (see preparation of silica gel below). Before gels can be prepared it is necessary to determine the quantity of N NaOH (Solution 1, Group A) required to adjust the pH to the desired level after the addition of NaHC03 and CaClz solutions (Solutions 5 and 6, Group A). M. Neutralize with N NaOH and note the amount added (x). 2 with N NaOH (Solution 1, Group A). Divide the sample (approximately 20 ml) into four aliquots and add varying amounts of the CaClz solution to each.
By means of a capillary pipette, a tuft of trichomes from a crude enrichment flask is transferred through several drops of steril tap water to remove most of the contaminating bacteria. 0. Standard sterilization techniques are used in all media preparation. A dry surface is obtained by allowing excess water to evaporate from the surface of the medium in a 30°C incubator for about 2 h with the lid of each dish propped open slightly. Between 4 and 6 h after inoculation each plate is examined under a dissecting microscope at 30 x magnification.