Improving the yield and quality of DNA isolated from white-rot fungi.
Identifieur interne : 000875 ( Main/Exploration ); précédent : 000874; suivant : 000876Improving the yield and quality of DNA isolated from white-rot fungi.
Auteurs : R C Kuhad [Inde] ; R K Kapoor ; R. LalSource :
- Folia microbiologica [ 0015-5632 ] ; 2004.
Descripteurs français
- KwdFr :
- ADN fongique (analyse), ADN fongique (isolement et purification), ADN fongique (métabolisme), Basidiomycota (composition chimique), Bromure de cétrimonium (MeSH), Chitinase (métabolisme), Composés de cétrimonium (composition chimique), DNA restriction enzymes (métabolisme), Détergents (MeSH), Mycologie (méthodes), Phanerochaete (composition chimique), Phénol (composition chimique), Pleurotus (composition chimique), Polyosides (composition chimique), Précipitation fractionnée (MeSH).
- MESH :
- analyse : ADN fongique.
- composition chimique : Basidiomycota, Composés de cétrimonium, Phanerochaete, Phénol, Pleurotus, Polyosides.
- isolement et purification : ADN fongique.
- métabolisme : ADN fongique, Chitinase, DNA restriction enzymes.
- méthodes : Mycologie.
- Bromure de cétrimonium, Détergents, Précipitation fractionnée.
English descriptors
- KwdEn :
- Basidiomycota (chemistry), Cetrimonium (MeSH), Cetrimonium Compounds (chemistry), Chitinases (metabolism), DNA Restriction Enzymes (metabolism), DNA, Fungal (analysis), DNA, Fungal (isolation & purification), DNA, Fungal (metabolism), Detergents (MeSH), Fractional Precipitation (MeSH), Mycology (methods), Phanerochaete (chemistry), Phenol (chemistry), Pleurotus (chemistry), Polysaccharides (chemistry).
- MESH :
- chemical , analysis : DNA, Fungal.
- chemical , chemistry : Cetrimonium Compounds, Phenol, Polysaccharides.
- chemical , isolation & purification : DNA, Fungal.
- chemical , metabolism : Chitinases, DNA Restriction Enzymes, DNA, Fungal.
- chemical : Cetrimonium, Detergents.
- chemistry : Basidiomycota, Phanerochaete, Pleurotus.
- methods : Mycology.
- Fractional Precipitation.
Abstract
A new simple method used to eliminate polysaccharides that cause problems during DNA isolation was established for 6 different white-rot fungi using 1% hexadecyltrimethylammonium bromide (CTAB) as wash buffer and followed by centrifugation. Variation in the DNA yield and quality was ascertained using precipitating agents, detergents and cell-wall-hydrolyzing chitinase. Considerable amount of exopolysaccharides from fungal biomass was removed with the use of 1% CTAB wash buffer followed by centrifugation. The DNA varied in terms of yield and quality. For the DNA extraction use of 2% SDS in extraction buffer worked best for Pycnoporus cinnabarinus, Cyathus bulleri, Cyathus striatus and Cyathus stercoreus, while 2% CTAB worked best for Phanerochaete chrysosporium and Pleurotus ostreatus. Elimination of phenol and use of absolute ethanol for precipitating DNA resulted in good yield and quality of DNA. This DNA was amenable to restriction endonuclease digestion.
DOI: 10.1007/BF02931383
PubMed: 15227780
Affiliations:
Links toward previous steps (curation, corpus...)
Le document en format XML
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<term>DNA Restriction Enzymes (metabolism)</term>
<term>DNA, Fungal (analysis)</term>
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<term>Bromure de cétrimonium (MeSH)</term>
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<term>DNA restriction enzymes (métabolisme)</term>
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<front><div type="abstract" xml:lang="en">A new simple method used to eliminate polysaccharides that cause problems during DNA isolation was established for 6 different white-rot fungi using 1% hexadecyltrimethylammonium bromide (CTAB) as wash buffer and followed by centrifugation. Variation in the DNA yield and quality was ascertained using precipitating agents, detergents and cell-wall-hydrolyzing chitinase. Considerable amount of exopolysaccharides from fungal biomass was removed with the use of 1% CTAB wash buffer followed by centrifugation. The DNA varied in terms of yield and quality. For the DNA extraction use of 2% SDS in extraction buffer worked best for Pycnoporus cinnabarinus, Cyathus bulleri, Cyathus striatus and Cyathus stercoreus, while 2% CTAB worked best for Phanerochaete chrysosporium and Pleurotus ostreatus. Elimination of phenol and use of absolute ethanol for precipitating DNA resulted in good yield and quality of DNA. This DNA was amenable to restriction endonuclease digestion.</div>
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<Abstract><AbstractText>A new simple method used to eliminate polysaccharides that cause problems during DNA isolation was established for 6 different white-rot fungi using 1% hexadecyltrimethylammonium bromide (CTAB) as wash buffer and followed by centrifugation. Variation in the DNA yield and quality was ascertained using precipitating agents, detergents and cell-wall-hydrolyzing chitinase. Considerable amount of exopolysaccharides from fungal biomass was removed with the use of 1% CTAB wash buffer followed by centrifugation. The DNA varied in terms of yield and quality. For the DNA extraction use of 2% SDS in extraction buffer worked best for Pycnoporus cinnabarinus, Cyathus bulleri, Cyathus striatus and Cyathus stercoreus, while 2% CTAB worked best for Phanerochaete chrysosporium and Pleurotus ostreatus. Elimination of phenol and use of absolute ethanol for precipitating DNA resulted in good yield and quality of DNA. This DNA was amenable to restriction endonuclease digestion.</AbstractText>
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