DGGE Fingerprinting of Bacterial Communities from very Humic Soils of South Siberia

Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F70883521%3A28110%2F02%3A63500556" target="_blank" >RIV/70883521:28110/02:63500556 - isvavai.cz</a>
Result on the web
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DOI - Digital Object Identifier
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Result language
angličtina
Original language name
DGGE Fingerprinting of Bacterial Communities from very Humic Soils of South Siberia
Original language description
To have a good chance of getting clearly visible band patterns on DGGE gel it is necessary to obtain good yield of PCR product which than can be loaded on a gel. It is a problem in case when the samples are humic soils. Fraction of humic substances display similar properties to DNA hence it can be uneasy to remove them during DNA isolation process and the substances consequently inhibit PCR. Additional cleanings of isolated DNA (Wizard DNA clean-up system, Promega) and subsequent tenfold dilution were sufficient and brought good PCR results. The most problematic sample needed three cleaning steps and tenfold dilution. The applicability of two approaches was tested: i) Direct use of Muyzer primers, when the PCR product can be readily loaded on DGGE gel;ii) PCR amplification of almost complete 16s rDNA sequence with use of primers homologous to very conservative terminal sequences, when the product was used as a template in next PCR with Muyzer primers. The nested approach appeared to be
Czech name
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Czech description
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Publication year
2002
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

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