Simple, Efficient, and Nondestructive DNA Extraction Protocol for Arthropods

Phillips, Aloysius; Simon, Chris

doi:10.1093/aesa/88.3.281

Cited by 73 publications

(57 citation statements)

References 3 publications

(3 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Specimens can also be dried from ethanol using chemical techniques. Phillips and Simon [39] presented a protocol for the isolation of DNA from pinned insects without destruction of the exoskeleton.…”

Section: Protocol 1 Methods Of Desiccationmentioning

confidence: 99%

Obtaining, Storing and Archiving Specimens and Tissue Samples for Use in Molecular Studies

Prendini¹,

Hanner²,

DeSalle³

2002

Techniques in Molecular Systematics and Evolution

View full text Add to dashboard Cite

Section: Protocol 1 Methods Of Desiccationmentioning

confidence: 99%

Obtaining, Storing and Archiving Specimens and Tissue Samples for Use in Molecular Studies

Prendini¹,

Hanner²,

DeSalle³

2002

Techniques in Molecular Systematics and Evolution

View full text Add to dashboard Cite

“…Genomic DNA from the bacterial isolates was extracted using the CTAB-DTAB method as described by Phillips and Simon (1995). First, each bacterium was inoculated in 5 mL sterile tryptic soy broth (TSB, Difco) and incubated overnight at 28 °C overnight following the method described by Kim and Jeong (2001).…”

Section: Dna Isolationmentioning

confidence: 99%

Identification of Elizabethkingia meningoseptica from American bullfrog (Rana catesbeiana) farmed in Sabah, Malaysia using PCR method and future management of outbreak

Ransangan¹,

Zainuri²,

Lal³

et al. 2013

MJM

View full text Add to dashboard Cite

Aims: High demand for frog meat in Malaysia especially the American bullfrog (Rana catesbeiana) has promoted intensive farming of the animal. However, the farming of American bullfrog is restricted by the occurrence of diseases. This study reports the first isolation of Elizabethkingia meningoseptica from specimens of American bullfrog that suffer from cataract and 'red-leg' syndrome. Methodology and Result: The pathogen was isolated from eyes and internal organs (liver, kidney and spleen) of the diseased bullfrog specimens. All the bacterial isolates were subjected to phenotypic characterization and antibiotic susceptibility assay, and further identified by using the 16S rDNA sequencing analysis. We designed two pair of specific PCR primers (22-25 mers) which are complimentary to the β-lactamase gene in the reference strain of E. meningoseptica ATCC49470. The result showed all the bacterial isolates shared similar phenotypic characters and antibiotic susceptibility. BLAST analysis of the 16S rDNA sequences indicated that the bacterial isolates had very high sequence homology (100%) with E. meningospetica ATCC49470 and E. meningoseptica isolates from mosquito. The two PCR primers were very specific to E. meningoseptica isolates of this study. Conclusion, significance and impact of study: This is the first isolation and characterization of bacterial pathogen, E. meningoseptica in cultured American bullfrog (Rana catesbeina) that suffered from eye cataract and 'red-leg' syndrome in Sabah, Malaysia. It is suspected that one of the possible transmission routes of the bacterial pathogen could be via mosquito bites. The findings suggest that there is urgent requirement for standard guideline of good farming practice to be adopted in frog farms throughout the country. Such a guideline can help in minimizing economic losses, preventing transmission of the zoonotic bacterial pathogen to farm workers, and sustaining the industry in Malaysia and upgrading frog meat quality for international market.

show abstract

“…Damselfly samples were frozen or preserved in 8% DTAB (dodecyltrimethylammonium bromide), 1.5 M NaCl, 0.05 M EDTA, and 0.10 M Tris/HCl, pH 8.6 (Phillips and Simon, 1995), or 0.25 M EDTA/2.5% SDS/0.5 M Tris, pH 9.2 (Cockburn and Fritz, 1996), or 70% isopropanol. DNA was extracted using a modification of Phillips and Simon's (1995) DTAB/CTAB extraction protocol detailed by Chippindale et al (1998).…”

Section: Taxa Sampledmentioning

confidence: 99%

“…DNA was extracted using a modification of Phillips and Simon's (1995) DTAB/CTAB extraction protocol detailed by Chippindale et al (1998). Collecting localities are listed in Table 1.…”

Section: Taxa Sampledmentioning

confidence: 99%

Phylogenetic Relationships of North American Damselflies of the GenusIschnura(Odonata: Zygoptera: Coenagrionidae) Based on Sequences of Three Mitochondrial Genes

Chippindale

Davé

Whitmore

et al. 1999

Molecular Phylogenetics and Evolution

View full text Add to dashboard Cite

Simple, Efficient, and Nondestructive DNA Extraction Protocol for Arthropods

Abstract: We present a DNA extraction technique that does not require the destruction of the insect exoskeleton. This technique is, therefore, of great utility for extracting from museum specimens DNA suitable for the polymerase chain reaction and subsequent DNA sequencing.

Cited by 73 publications

References 3 publications

Obtaining, Storing and Archiving Specimens and Tissue Samples for Use in Molecular Studies

Obtaining, Storing and Archiving Specimens and Tissue Samples for Use in Molecular Studies

Identification of Elizabethkingia meningoseptica from American bullfrog (Rana catesbeiana) farmed in Sabah, Malaysia using PCR method and future management of outbreak

Phylogenetic Relationships of North American Damselflies of the GenusIschnura(Odonata: Zygoptera: Coenagrionidae) Based on Sequences of Three Mitochondrial Genes

Contact Info

Product

Resources

About