Journal
JOURNAL OF MICROBIOLOGICAL METHODS
Volume 70, Issue 3, Pages 565-569Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.mimet.2007.06.013
Keywords
DGGE; T-RFLP; ARISA; ruminant; profiling; diversity; cytophage-flexibacter-bactcroides; community
Categories
Funding
- Biotechnology and Biological Sciences Research Council [BBS/E/G/00003008] Funding Source: researchfish
- Biotechnology and Biological Sciences Research Council [BBS/E/G/00003008] Funding Source: Medline
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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.
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