Specificity and sensitivity of eubacterial primers utilized for molecular profiling of bacteria within complex microbial ecosystems
Specificity and sensitivity of eubacterial primers utilized for molecular profiling of bacteria within complex microbial ecosystems
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DOI：
10.1016/j.mimet.2007.06.013
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发表时间：
2007-09-01
影响因子：
2.2
通讯作者：
Scollan, N. D.
中科院分区：
文献类型：
--
作者：
Huws, S. A.;Edwards, J. E.;Scollan, N. D.
在复杂群落中对真细菌多样性进行有效分析需要对真细菌16S rRNA具有特异性的引物。已发表的引物对立方细菌和古细菌16S rRNA以及原生动物和真菌18S rRNA的特异性进行了计算机评价。随后用瘤胃样品验证了V3和V6-V8 （F968gc和R1401）变性梯度凝胶电泳（DGGE）引物的特异性和敏感性。并对采用触地PCR循环条件的效果进行了评价。对于瘤胃样品中真菌体的DGGE分析，在不使用触地PCR的情况下，引物F968gc和r1401被证明是最特异和最敏感的。(C) 2007 Elsevier B.V.版权所有
Efficient profiling of eubacterial diversity within complex communities requires that primers are specific for eubacterial 16S rRNA. Specificity of published primers against cubacterial and archaeal 16S rRNA as well as protozoal and fungal 18S rRNA was assessed in silico. The specificity and sensitivity of the V3 and V6-V8 (F968gc and R1401) Denaturing Gradient Gel Electrophoresis (DGGE) primers was subsequently verified using rumen-derived samples. An assessment of the effects of employing touchdown PCR cycling conditions was also made. For DGGE profiling of eubacteria within rumen samples, primers F968gc and R 1401 proved the most specific and sensitive providing that touchdown PCR is not used. (C) 2007 Elsevier B.V. All rights reserved.