BIOLOG及.ppt

Garlan和Mills1991年提出群落水平生理学指纹方法（CLPP）是通过检测微生物样品对底物利用模式来反映种群组成的酶活性分析方法 具体而言，通过检测微生物样品对多种不同的单一碳源基质利用能力，来确定哪些基质可以作为能源，从而产生对基质利用的生理代谢指纹 方法流程 水溶液制备（过滤或振荡提取） 水溶液稀释 加入微孔板 培养 590nm波长测定吸光度 95个孔吸光度的平均值（average well color development, AWCD） 影响平板微孔显色的因素较多，除了培养液中微生物的种群组成、数量与活性外，样品的预处理，培 养时间和温度，培养液含有干扰物质等因素都会影响平板显色结果 使用biolog分析更多的是对环境微生物群落进行比较和识别，以及进行群落活性与功能的分析。还 不能直接通过Biolog代谢指纹分析微生物群落结构信息。 仅能鉴定快速生长的微生物；测试盘内近中性的缓冲体系、高浓度的碳源；指示剂红四氮唑（TTC）对微生物有毒 方法的不足 * * * * * Using a Streakerz, pick up the growth from the outer half of one of the legs of growth. ? Be careful not to pick up any of the agar media since this could introduce another carbon source. ? Apply the growth, on the tip of the stick, to the inner wall of a glass tube. ? A dry, sterile, 20 x 150 mm tube is used for this step. ? Using a circular, up and down motion, with the Streakertz, work the organism mass against the inner surface of the tube. ? Harvest more growth from the second leg and add this to the homogenized bacteria in the tube. ? Break up the mass by continuing the same blending motion in the dry tube as before. ? The same technique is used to collect the growth on the third and fourth legs of the plus. ? The complete process of harvesting the growth from all four legs of the plus, and the homogenization of this growth against the inside of the dry tube is being shown. This is being done to illustrate the amount of time and effort that is required for this procedure. ? Following this procedure in detail will make the difference between a successful outcome and an indeterminate result. ? The inner surface of the tube should look like it was coated with an evenly distributed, thin, pasty film ? The purpose of the dry tube technique is to completely break up the bacterial aggregates. ? The growth at the tips of the plus is metabolically more active than the interior of the plus. Organisms from the outer area will give optimal results in the MicroPlate. ? The action of pressing the bacterial g