Specificity and sensitivity of eubacterial primers utilized for molecular profiling of bacteria within complex microbial ecosystems

被引：125

作者：

Huws, S. A. ^{[1
]}

Edwards, J. E. ^{[1
]}

Kim, E. J. ^{[1
]}

Scollan, N. D. ^{[1
]}

机构：

[1] Inst Grassland & Environm Res, Aberystwyth SY23 3EB, Dyfed, Wales

来源：

JOURNAL OF MICROBIOLOGICAL METHODS | 2007年 / 70卷 / 03期

基金：

英国生物技术与生命科学研究理事会;

关键词：

DGGE; T-RFLP; ARISA; ruminant; profiling; diversity; cytophage-flexibacter-bactcroides; community;

D O I：

10.1016/j.mimet.2007.06.013

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页码：565 / 569

页数：5

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