A simple and reliable method for DNA extraction from bivalve mantle

Aranishi, Futoshi; Okimoto, Takane

doi:10.1007/bf03194632

Cited by 24 publications

(22 citation statements)

References 13 publications

(5 reference statements)

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“…However, we found that the modified chelex-100 protocol was the best method to extract DNA from stool. Although there are methodological differences between our study and others [32,33] , the conclusion is the same that this method is simple and efficient for PCR.…”

Section: Discussionsupporting

confidence: 45%

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A simple and rapid method for extracting bacterial DNA from intestinal microflora for ERIC-PCR detection

Yang¹,

Wang²,

Cheng³

et al. 2008

WJG

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AIM:To develop a simple and convenient method for extracting genomic DNA from intestinal microflora for enterobacterial repetitive intergenic consensus (ERIC)-PCR detection. METHODS:Five methods of extracting bacterial DNA, including Tris-EDTA buffer, chelex-100, ultrapure water, 2% sodium dodecyl sulfate and 10% Triton-100 with and without sonication, were compared with the commercial fecal DNA extraction kit method, which is considered as the gold standard for DNA extraction. The comparison was based on the yield and purity of DNA and the indexes of the structure and property of micro-organisms that were reflected by ERIC-PCR. RESULTS:The yield and purity of DNA obtained by the chelex method was similar to that obtained with the fecal DNA kit. The ERIC-PCR results obtained for the DNA extracted by the chelex method and those obtained for DNA extracted with the fecal DNA kit were basically the same. CONCLUSION:The chelex method is recommended for ERIC-PCR experiments in view of its simplicity and costeffectiveness; and it is suitable for extracting total DNA from intestinal micro-organisms, particularly for handling a large number of samples.

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Section: Discussionsupporting

confidence: 45%

“…The results vary by different methods [32,33] . The use of chelex-100 has been recommended for DNA extraction in some papers, but other reports have not regarded it as being optimum because of its lowest efficiency for DNA amplification [32] .…”

Section: Discussionmentioning

confidence: 99%

A simple and rapid method for extracting bacterial DNA from intestinal microflora for ERIC-PCR detection

Yang¹,

Wang²,

Cheng³

et al. 2008

WJG

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“…Deoxyribonucleic acid (DNA) extraction-Ctenophores collected at each station (Table 1) were placed individually or in groups of up to five individuals (those #1 mm) into Eppendorf tubes containing 400 mL of 4 mol L 21 urea, 1% Tween 20, 1% Nonidet P-40, 10% Chelex 100, and 0.005 mg proteinase K in sterile water (Aranishi and Okimoto 2006) within 2 h after collection. The tubes were incubated at 55uC for 20 min, and then at 105uC for 8 min.…”

Section: Methodsmentioning

confidence: 99%

Molecular evidence for the occurrence of ctenophore Mertensia ovum in the northern Baltic Sea and implications for the status of the Mnemiopsis leidyi invasion

Gorokhova

Lehtiniemi²,

Viitasalo-Frösen³

et al. 2009

Limnology & Oceanography

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Nucleotide sequence analysis of 18S ribosomal RNA gene (rRNA), internal transcribed spacer, and 5.8S rRNA was used for taxonomic identification of ctenophores collected in the northern Baltic Sea, where invasive Mnemiopsis leidyi and native Pleurobrachia pileus have been reported to occur. Contrary to previous reports, sequence analysis of 53 randomly selected specimens from seven stations revealed that none of them were M. leidyi or P. pileus. The 18S rRNA and 5.8S rRNA sequences were 100% identical to those of Mertensia ovum, a ctenophore with a broad Arctic and circumboreal distribution, which has never been reported to occur in the Baltic Sea. Polymerase chain reaction screening with primers designed to amplify all three species, and using ctenophores collected by vertically stratified sampling, confirmed that all ctenophores collected in this survey were M. ovum. The ctenophore abundance was high, up to 4500 individuals m 22 , positively correlating with salinity. Our findings emphasize the utility of applying molecular tools to biological surveys and the importance of rigorous species identification. They also indicate that M. leidyi, which is a threat to the southern Baltic ecosystem, does not occur in the northern part of the sea, and call for a pan-Baltic survey to establish current distributions of ctenophores, both native and invasive.

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“…In recent decades, researchers have been increasingly more likely to solve taxonomic problems in certain species using polymorphic genetic markers, also widely used in phylogenetic studies and population genetics (Yue and Orban, 2001;Aranishi and Okimoto, 2006). One of the most used genes is mitochondrial cytochrome oxidase I (COI), which has high phylogenetic signal and rapid evolutionary rate, which is important for characterizing populations, subspecies and species (Harrison, 1989).…”

Section: Introductionmentioning

confidence: 99%

Novel DNA extraction assay for molecular identification of Aedes spp eggs

Freitas¹,

Gomes-Júnior²,

Batista³

et al. 2014

Genet. Mol. Res.

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ABSTRACT.Aedes aegypti and A. albopictus represent the two most important species of mosquitoes in relation to dengue virus transmission both in the Americas and Asia. However, the study of theses species generally requires the establishment of a colony for the larvae to hatch, or waiting for the adult development to perform its taxonomic classification, which is time consuming. Thus, the establishment of new methods aimed at obtaining DNA directly from the mosquito eggs is relevant. Accordingly, we compared a new approach based on Chelex ® 100 resin with the standard STE method to extract DNA from the eggs of Aedes spp to molecularly identify these vectors. The Chelex ® 100 resin approach was very efficient, as satisfactory amounts of DNA were obtained, making it possible to amplify and sequence a mitochondrial DNA barcode region widely used to identify species. The STE protocol yielded substantial amounts of DNA, but the 260/280 optical density ratio indicated a low quality, precluding amplification. This new method proved quite effective in obtaining DNA from even a single mosquito egg, and it can thus be applied in population genetic studies of various vector insects to enhance monitoring programs.

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A simple and reliable method for DNA extraction from bivalve mantle

Cited by 24 publications

References 13 publications

A simple and rapid method for extracting bacterial DNA from intestinal microflora for ERIC-PCR detection

A simple and rapid method for extracting bacterial DNA from intestinal microflora for ERIC-PCR detection

Molecular evidence for the occurrence of ctenophore Mertensia ovum in the northern Baltic Sea and implications for the status of the Mnemiopsis leidyi invasion

Novel DNA extraction assay for molecular identification of Aedes spp eggs

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