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Because an early clue about the underlying genus or species may have a major impact on patient management and antimicrobial therapy, the eagerly awaited results of microscopy and Gram staining might be complemented by the nearly isochronous but more discriminative results of eubacterial PCR and high-resolution melting.
The sequencing of the 16S rDNA allowed a distinction among different bacterial species on the basis of the hypervariable region, which is flanked from conserved eubacterial sequences.
One primer pair (341f-GC and 907r) was used for general detection of bacteria by amplification of a 550-bp fragment from the region V3 of eubacterial 16S rRNA (Muyzer et al.
Standard PCR was performed by using the eubacterial broad-range 16S rDNA primer set 536F: 5'-CAGCAGCCGCGGTAATAC-3' and rp2: 5'-ACGGC TACCTTGTTACGACTT-3' (Eurogentec, Seraing, Belgium).
Our universal eubacterial 16S rDNA approach used analysis of small clone libraries from each animal to detect those agents representing [greater than or equal to] 10% of the 16S operons in lung tissue with high (>95%) confidence.
The diversity of the eubacterial communities in these mats was determined using culture-independent community DNA.
The V3 region of eubacterial rrs genes was amplified using primers f5'-GC-clamp-ACGGGYGCAGCAGGCG CGA-3' and r5'-GWATTACCGCGGCKGCTG-3' (Bano et al.
In order to assess the presence of other bacteria, primers for 16S eubacterial DNA for (27F 5'-AGAGTTTGATCMTGGCTCAG-3' and 1513R 5'- ACGGYTACCTTGTTACGACTT-3', ex Weis burg et al.
Eubacterial, archaebacterial, and eucaryotic genes that encode leaderless mRNA.
Under our hypothesis, the cytoskeleton results, like many other features, from the genetic complexity conferred by the forced integration of eubacterial genes into archaebacterial chromosomes," he says.
We amplified each sample by using the eubacterial universal 16S primer set 27F/338R described by Fierer et al.
The 16S rRNA genes from these isolates were PCR-amplified using the eubacterial primers 8F (5'-AGAGTTTGATCCTGGCTCAG-3') and 1492R (5'-GGTTACCTTGTTACGACYT-3') with the reaction mixture and PCR conditions described in Lundgren and Lehman (2010).