Rapid isolation of fungal genomic DNA suitable for long distance PCR

Maeseneire, Sofie L. De; Bogaert, Inge Van; Dauvrin, T.; Soetaert, Wim; Vandamme, Erick

doi:10.1007/s10529-007-9483-6

Cited by 10 publications

(1 citation statement)

References 23 publications

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“…The purity of the DNA was assessed with spectrophotometry via calculating A 260 /A 230 and A 260 /A 280 ratio, coextracted humic acids, phenol and other aromatic compounds absorb at 230 nm whereas DNA absorbs at 260 nm and protein at 280 nm, the high A 260 /A 230 and A 260 /A 280 ratio were indicative of purity of DNA [2,22]. The yield rates of DNA were estimated by the ratio of actual and theoretical values of DNA yields.…”

Section: Detection Methodsmentioning

confidence: 99%

A developed DNA extraction method for different soil samples

Liu

Yan

et al. 2010

J. Basic Microbiol.

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Four DNA extraction methods namely SDS-hyperhaline method (I), modified SDS-hyperhaline method (II), indirect method (III), alkaline lysis method (IV) were evaluated by comparing DNA yield, spectrophotometric quality, genomic integrity and PCR suitability in this paper. The results showed that high DNA yields were obtained by method I, II and IV. However, higher quality of DNA was gained by method III and IV. Based on the results of the Pulsed-Field Gel Electrophoresis (PFGE), the completeness of DNA extracted by method IV was the best. About 6.0 microg DNA can be recovered from 1.0 g soil by method IV which involved to lysis cell by SDS and to precipitate impurities by adding potassium acetate and magnesium chloride Therefore, it is confirmed that method IV is a novel, reliable and versatile method for large-scale DNA extraction involving less purification steps for various soil samples.

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Section: Detection Methodsmentioning

confidence: 99%

A developed DNA extraction method for different soil samples

Liu

Yan

et al. 2010

J. Basic Microbiol.

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Drill-assisted genomic DNA extraction from Botrytis cinerea

et al. 2008

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Current DNA extraction protocols for genomic DNA from Botrytis cinerea almost always start with mycelium that has been reduced to powder with liquid N(2) in a mortar, and this makes their application to a large number of samples slow and cumbersome. Here we present an adaptation of an existing method [Möller et al. (1992) Nucleic Acids Res 20: 6115-6116] for which the initial steps have been modified, including the homogenization of the fungus with sand and the aid of a common household drill. This method allows the processing of large number of samples in much shorter times and generates an average of 4 mug DNA per sample, of sufficient quality for use in PCR and Southern blotting.

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Development of a Simple and Cost-Effective Bead-Milling Method for DNA Extraction from Fish Muscles

et al. 2014

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In the fish food sector, due to a growing globalization of the market, where intentional and unintentional frauds reach alarming levels, the molecular analysis is increasingly used by both Official agencies, to enforce the law on traceability, and private companies, to verify the quality of goods. DNA extraction represents a necessary and critical step for all types of DNA analysis. Among the drawbacks associated with this procedure, there are handling of toxic materials, low DNA yield and low throughput, due to time-consuming manual procedures. In this work, to overcome some of these problems, we developed an alternative method based on a bead milling procedure without proteinase K digestion. The new method was then compared with both a salting out protocol, developed in a previous work, and a commercial kit. Yield, spectrophotometric purity, electrophoretic degradation pattern, and amplificability of the extracted DNA were assessed. In particular, DNA amplificability was evaluated by comparing the band intensity on the gel, after amplification of the 16S rRNA and COI genes with a conventional PCR, and the takeoff cycles, after amplification of the 16S rRNA gene with a real-time PCR. The results showed that the bead-based method allowed to obtain acceptable amounts of DNA, with good purity and good characteristics of amplificability. Although the salting out method remains the most effective protocol in terms of pure performances, the bead-milling procedure can be considered a valid alternative, in the light of its lower demand in terms of labor and costs.

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Rapid isolation of fungal genomic DNA suitable for long distance PCR

Cited by 10 publications

References 23 publications

A developed DNA extraction method for different soil samples

A developed DNA extraction method for different soil samples

Drill-assisted genomic DNA extraction from Botrytis cinerea

Development of a Simple and Cost-Effective Bead-Milling Method for DNA Extraction from Fish Muscles

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