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

Kód výsledku v IS VaVaI

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Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

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

Popis výsledku v původním jazyce

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

Název v anglickém jazyce

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

Popis výsledku anglicky

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

Druh

D - Stať ve sborníku

CEP obor

EB - Genetika a molekulární biologie

OECD FORD obor

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Projekt

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Návaznosti

Z - Vyzkumny zamer (s odkazem do CEZ)

Rok uplatnění

2002

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název statě ve sborníku

XVIII. biochemický zjazd - zborník

ISBN

nemá

ISSN

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e-ISSN

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Počet stran výsledku

1

Strana od-do

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Název nakladatele

Slovenská spoločnosť pre biochémiu a mol.biológiu pri SAV

Místo vydání

Bratislava

Místo konání akce

Stará Lesná

Datum konání akce

10. 9. 2002

Typ akce podle státní příslušnosti

EUR - Evropská akce

Kód UT WoS článku

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