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Optimization of the DGGE band identification method
Kušar, Darja (Avtor), Avguštin, Gorazd (Avtor)

URLURL - Predstavitvena datoteka, za dostop obiščite http://www.springerlink.com/content/a48w624573471112/fulltext.pdf Povezava se odpre v novem oknu

Izvleček
Denaturant gradient gel electrophoresis (DGGE) enables insight into the diversity of the studied microbial communities on the basis of separation of PCR amplification products according to their nucelotide sequence composition. However, the success of the method is accompanied by the inherent appearance of various sequence artifacts that bias the impression of community structure by generating additional bands representing no virtual microbes. PCR-DGGE artifacts require optimization of the method when aiming at the phylogenetic identification of the selected DGGE bands. The aim of our study was to develop a procedure which will increase the reliability of the identification. Samples of rumen fluid were used for the optimization since they contain a complex microbial community that supports the generation of artifactual bands. An optimized procedure following band excision and elution of microbial DNA is proposed including nuclease treatment, selection of DNA polymerase with proofreading activity, and cloning prior to sequencing and identification analysis.

Jezik:Angleški jezik
Ključne besede:mikrobiologija, metode, elektroforeza, DGGE
Vrsta gradiva:Delo ni kategorizirano (r6)
Tipologija:1.01 - Izvirni znanstveni članek
Organizacija:BF - Biotehniška fakulteta
Leto izida:2012
Št. strani:301-306
Številčenje:Vol. 57, no. 4
UDK:579
ISSN pri članku:0015-5632
DOI:10.1007/s12223-012-0130-9 Povezava se odpre v novem oknu
COBISS.SI-ID:3042696 Povezava se odpre v novem oknu
Število ogledov:937
Število prenosov:142
Metapodatki:XML RDF-CHPDL DC-XML DC-RDF
 
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Gradivo je del revije

Naslov:Folia microbiologica
Skrajšan naslov:Folia microbiol.
Založnik:Springer
ISSN:0015-5632
COBISS.SI-ID:25470976 Povezava se odpre v novem oknu

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