Metagenomic dna isolation from sheep feces and pcr detection of several rumen bacteria
<b>Aim:</b> The aim of this study was to isolate metagenomic DNA from fresh and dry sheep feces and to detect several rumen bacteria using the specific primers.<p> <b>Materials and Methods:</b> The metagenomic DNA isolation was performed by using commercial I-Genomic St...
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| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Selcuk University Press
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| Series: | Eurasian Journal of Veterinary Sciences |
| Subjects: | |
| Online Access: | http://eurasianjvetsci.org/pdf.php3?id=1110 |
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| Summary: | <b>Aim:</b> The aim of this study was to isolate metagenomic DNA
from fresh and dry sheep feces and to detect several rumen
bacteria using the specific primers.<p>
<b>Materials and Methods:</b> The metagenomic DNA isolation
was performed by using commercial I-Genomic Stool DNA
Isolation Kit. Anaerovibrio lipolytica, Fibrobacter succinogenes,
Prevotella bryantii, Prevotella ruminicola, Ruminobacter
amylophilus, Ruminococcus albus, Ruminococcus flavefaciens,
Streptococcus bovis, Selenomonas ruminantium and Succinovibrio
dextrinosolvens were screened using metagenomic
DNA with polymerase chain reaction and spesific primers.
Reaction of 16S rRNA region with SphI was carried out to
confirm the absence of R. amylophilus, R. albus and S. dextrinosolvens.<p>
<b>Results:</b> Fecal samples dried rapidly and lost its 53.72% of
fresh mass. After the DNA isolations from fresh and dried
samples, DNA concentrations and purity were varied between
25.60-59.50 ng/μL and 1.72-1.90, respectively. It was observed
that fecal inhibitors had no effect on PCR. A. lipolytica,
F. succinogenes, P. bryantii, P. ruminicola, R. flavefaciens, S. bovis
and S. ruminantium were detected with specific primers
however PCR did not reveal the presence of R. amylophilus,
R. albus and S. dextrinosolvens. SphI digestion of 16S rDNA
regions has confirmed this result.<p>
<b>Conclusions:</b> In this study, effect of drying in natural conditions
on metagenomic DNA isolation from fecal samples was
determined. Furthermore, PCR detection of several rumen
bacteria was performed by using isolated DNA. In conclusion,
it may be stated that the metagenomic DNA isolated from
dried fecal samples could be an effective tool for the detection
of bacterial populations. |
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| ISSN: | 1309-6958 2146-1953 |