By Christopher L. Clayton, Harry L. T. Mobley
Helicobacter pylori Protocols bargains a superb number of cutting-edge protocols for the identity and molecular manipulation of H. pylori. The authoritative individuals provide specific and without problems reproducible protocols for the culturing of H. pylori, for the isolation and restrict endonuclease digestion of H. pylori chromosomal DNA, and for the transformation and insertional mutagenesis of H. pylori. in addition they offer molecular epidemiological innovations, together with ribotyping, PCR-RFLP, and RAPD-PCR. those systems were built by way of top practitioners to resolve the tough technical difficulties created via the applying of the strong bacterial genetic and molecular cloning ideas to H. pylori.
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Extra info for Helicobacter pylori Protocols
2. Materials 1. The strainsthat have been grown m the defined medium are: NCTC 11637 (type strain), Roberts (clinical isolate from Manchester, UK), and 8 isolates from stomach biopsies collected during 199 l-l 992 m Birmingham, UK Two other clmical isolates failed to grow m the medium, but these also grew poorly in standard media. 2~pm syrmge filters [Gelman Sciences, Northampton, UK or Sartonus, depending on volume]), and store at 4°C for up to 6 mo (see Note 1). 5% (w/v) m water and sterilized by membrane filtration.
139,3023-3028. 7. , and Nilsson, L. E. (1993) Morphologic conversion of Helicobacter pylorl from bacdlary form to coccoid form evaluated by biolumtnescence, microscopy and viable count. Acta Gastro-Enterol Belg. Suppl 56,52. 8 Catrenich, C. E and Makin, K. M (1991) Charactertzatlon of the morphologic conversion of Helicobacter pylorl from bacillary to coccotd form. &and. J Gastroenterol 26(Suppl. Ml), 58-64. 9. , and Dainelli, B. (1993) Morphological forms in Helzcobacterpylorl Acta Gastro-Enter01 Belg Suppl 56, 108.
45, and 5 pm pore size Fluorescein diacetate (FDA). 96-prong moculator for 96-well microtiter trays (Denley). 3. 1. Minimum Inhibitory Concentration (M/C) 1. Prepare cultures by subculturmg H pylon onto freshly poured chocolate Columbia agar plates and incubate for 48 h mtcroaerobically (see Note 1) 2. Prepare doubling dilutions of test compounds and add 1 mL to triple-vented agar plates Take into account l/20 dilution after addition of agar 3. Add 19-mL chocolate Columbia agar to each plate and ensure the test compounds are suitably mixed mto the agar.