Sequencing and analysis: Sequencing of the PCR product of 16S rDNA
and blaNDM-1 genes of the representative samples was carried out commercially through Macrogen, South Korea.
An amplified fragment of 222bp from 16S rDNA
, 424bp from XCF/XCR and 500bp from XACF/XACR was observed from all collected citrus cultivars.
haemolytica was confirmed by complete sequencing of the 16S rDNA
gene with 99% identity in all analyzed isolates.
Specific primers for 16S rDNA
of full-length 1.6kb gene were used (Bukhari and Shakoori, 2010).
About 425 bp 16S rDNA
fragments were purified using QIAquick PCR purification kit (QIAGEN, Valencia, CA, USA) according to the manufacturer's instructions and sequenced with the same primers using the sequencer (Gene analyzer 3121) by Macrogen co., Korea.
If identification was still incorrect following the proteic extraction protocol, we performed 16S rDNA
sequencing systematically, as previously described (16), in 3 situations: when the identification score was <1.9 despite proteic extraction, when multiple different species were recognized with a correct identification score, and when a bacterium was isolated for the first time in the clinical microbiology laboratory.
Genetik analiz icin 16S+1 (5'-CTGCTCAATGATTTTTTAAATTGCTGTGG-3') ve 16S-1 (5'-CCGGTCTGAACTCAGATCAAGT-3') primerleri kullanilarak ixodid kenelerin 16S ribozomal DNA (16S rDNA
) bolgesinin yaklasik 460 baz cifti (basepair = bp) uzunlugunda fragmentini amplifiye eden PZR protokolu uygulanmistir (14).
Out of 270 bacterial isolates obtained from banknotes, 61 were subjected to 16S rDNA
amplification using universal primers.
Comparison of broad range 16S rDNA
PCR and conventional blood culture for diagnosis of sepsis in the newborn: a case control study.
First, they selected a set of universal primers targeting a conserved region in the 16S rDNA
of the bacterial genome.
Quantitative PCR was used to test for the presence of and to determine the amount of MeS 16S rDNA
relative to total bacterial 16S rDNA
present in DNA isolated from Mytilus edulis tissue samples.
The 16S rDNA
sequences were aligned with 16S rRNA sequences from GenBank to identify organisms using BLAST analysis .