16S核糖体RNA
细菌
牛血清白蛋白
特里斯
色谱法
聚合酶链反应
微生物
化学
核糖体RNA
悬挂(拓扑)
分子生物学
氨基酸
微生物学
生物
生物化学
基因
遗传学
数学
同伦
纯数学
作者
D. McGregor,S. N. Forster,John Steven,John R. Adair,Sophie E.C. Leary,Dario L. Leslie,William J. Harris,R.W. Titball
出处
期刊:BioTechniques
[Future Science Ltd]
日期:1996-09-01
卷期号:21 (3): 463-471
被引量:47
摘要
The effect of buffer composition on simultaneous PCR amplification of 16S rRNA gene fragments of five bacterial species was examined using a number of different buffer systems. Tris-based PCR buffers at final concentrations of 10 mM proved unreliable. However, when the final concentration of Tris was increased to 75 mM, all five samples were routinely detected. The use of other buffers, 3-[(1,1-dimethyl-2-hydroxyethyl) amino]-2-hydroxypropanesulfonic acid (AMPSO) and 3-[cyclohexylamino]-2-hydroxy-1-propanesulfonic acid (CAPSO), resulted in PCR amplification of five products even at low final concentrations (10 mM). The presence of certain proteins in the amplification reaction could overcome an inhibitory effect seen when soil suspension was present in the reaction, as might occur when testing field samples for the presence of bacteria. Bovine serum albumin was found to be the most effective additive tested in overcoming inhibition.
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