Kreader C A
Microbiology Research Division, Environmental Monitoring Systems Laboratory, U.S. Environmental Protection Agency, Cincinnati, Ohio 45268, USA.
Appl Environ Microbiol. 1996 Mar;62(3):1102-6. doi: 10.1128/aem.62.3.1102-1106.1996.
The benefits of adding bovine serum albumin (BSA) or T4 gene 32 protein (gp32) to PCR were evaluated with reaction mixtures containing substances that inhibit amplification. Whereas 10- to 1,000-fold more FeCl3, hemin, fulvic acids, humic acids, tannic acids, or extracts from feces, freshwater, or marine water were accommodated in PCR when either 400 ng of BSA per microl or 150 ng of gp32 per microl was included in the reactions, neither BSA nor gp32 relieved interference significantly when minimum inhibitory levels of bile salts, bilirubin, EDTA, NaCl, sodium dodecyl sulfate, or Triton X-100 were present. Use of BSA and gp32 together offered no more relief of inhibition than either alone at its optimal level, and neither protein had any noticeable effect on amplification in the absence of inhibitors.
在含有抑制扩增物质的反应混合物中评估了向聚合酶链反应（PCR）中添加牛血清白蛋白（BSA）或T4基因32蛋白（gp32）的益处。当反应中每微升包含400 ng BSA或每微升150 ng gp32时，PCR能够容纳比原来多10至1000倍的氯化铁、血红素、富里酸、腐殖酸、单宁酸或来自粪便、淡水或海水中的提取物；然而，当存在胆汁盐、胆红素、乙二胺四乙酸（EDTA）、氯化钠、十二烷基硫酸钠或聚山梨醇酯80（Triton X - 100）的最小抑制水平时，BSA和gp32均不能显著减轻干扰。在最佳水平下，同时使用BSA和gp32与单独使用其中任何一种相比，对抑制的缓解作用并无更大效果，并且在没有抑制剂的情况下，这两种蛋白质对扩增均无明显影响。
