Detection of Genetically Modified Crops and Their Derivatives: Critical Steps in Sample Preparation and Extraction

Terry, Catherine; Harris, Neil; Parkes, Helen C.

doi:10.1093/jaoac/85.3.768

Cited by 64 publications

(21 citation statements)

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“…Although it has been reported that concentrations of NaCl >25 mM inhibit PCR [19], our PCR analyses were successful despite the very high concentrations of NaCl used. Since washing the DNA pellet with ethanol removes NaCl [20], multiple washes with ethanol greatly reduce the concentration of NaCl. High concentration of EDTA prevented DNA degradation.…”

Section: Discussionmentioning

confidence: 99%

A Simple and Efficient Method for Extracting S. Rolfsii DNA For PCR Based Diversity Studies

As¹,

Kato²,

Cm³

2018

J Plant Pathol Microbiol

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Section: Discussionmentioning

confidence: 99%

A Simple and Efficient Method for Extracting S. Rolfsii DNA For PCR Based Diversity Studies

As¹,

Kato²,

Cm³

2018

J Plant Pathol Microbiol

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“…Cream is mainly composed of lipids such as oils or waxes [34,35]. Hexane or chloroform treatment minimizes PCR inhibition during DNA extraction from lipid-rich cosmetics [36,37]. However, the use of organic solvents in the extraction process is tedious and cumbersome; thus, most commercial kits do not use organic solvents.…”

Section: Detection Limit Of the Real-time Pcr Assaymentioning

confidence: 99%

Effect of DNA extraction methods on the detection of porcine ingredients in halal cosmetics using real-time PCR

Kim

Lee

et al. 2018

Appl Biol Chem

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In recent years, halal cosmetics have attracted considerable attention worldwide. We developed a realtime PCR assay based on the mitochondrial gene ndh5 for rapid detection of porcine ingredients in halal cosmetic products. We also compared several DNA extraction methods for the most efficient approach in different types of cosmetics. Porcine template DNA was spiked into three types of cosmetics (liquid-type and powder-type mask packs, and cream) and extracted with five commercial DNA extraction kits and the CTAB method. The extraction efficiency of each method was evaluated by determining the detection limits of real-time PCR assay. The lowest detection limit of real-time PCR for each cosmetic product was as follows: 2.28 9 10 0 copies for liquid-type mask pack when the Power Prep TM DNA extraction kit and TIANamp Genomic DNA kit were used, 2.28 9 10 1 copies for powder-type mask pack when QIAamp DNA stool mini kit and the Power Prep TM DNA extraction kit were used, and 2.28 9 10 0 copies for cream when the Power Prep TM DNA extraction kit was used. The pig-specific real-time PCR assay facilitated the detection of trace amounts of the template DNA in cosmetics, and an appropriate DNA extraction method was used depending on the type of cosmetics.

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“…To overcome this problem, the cetyl trimethylammonium bromide (CTAB) method can be applied with the addition of particular chemical and enzymatic treatments (Terry et al, 2002). To remove polysaccharides and proteins, treatments with enzymes such as pectinase, cellulase, hemicellulose, a-amylase, proteinase K and glycoside hydrolases can be used (Rether et al, 1993;Demeke & Jenkins, 2010).…”

Section: Molecular Fungal Detection Assaysmentioning

confidence: 99%

Diagnostic methods for detecting fungal pathogens on vegetable seeds

2016

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Early diagnosis of seedborne fungal pathogens is particularly important, as often, infected seeds appear symptomless; seed diagnosis can avoid uncontrolled propagation of pathogens through long‐distance exchange of such material. This will prevent economic losses and unnecessary use of fungicides, so reducing costs and the introduction of toxic substances into the environment. Traditional techniques for detection of seedborne fungi are based on incubation and grow‐out methods. Although these are frequently used because of their simplicity of application, they are time‐consuming, require mycological skills, and are sometimes not sensitive enough to low levels of seed infection. Recently, new identification techniques, based on DNA analysis, have been applied and are very efficient due to high sensitivity and specificity. The most common technique is conventional PCR, while other recent techniques include nested PCR, to obviate low levels of target pathogens, multiplex PCR, to detect several pathogens simultaneously, real‐time PCR, to quantify fungi on seeds, and magnetic‐capture hybridization PCR. The main drawbacks of molecular methods are the inability to distinguish between vital and non‐vital inocula, and the difficulty in obtaining quality DNA template, due to PCR inhibitors in seeds. To reduce inhibitory effects, several modified PCR protocols, such as loop‐mediated isothermal amplification, and non‐destructive testing methods have been developed. Loop‐mediated isothermal amplification and next‐generation sequencing have been widely applied in nucleic acid analysis and, given the numerous advantages provided, their application can be substantially extended in the future for detection of fungal pathogens in seeds.

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Detection of Genetically Modified Crops and Their Derivatives: Critical Steps in Sample Preparation and Extraction

Cited by 64 publications

References 0 publications

A Simple and Efficient Method for Extracting S. Rolfsii DNA For PCR Based Diversity Studies

A Simple and Efficient Method for Extracting S. Rolfsii DNA For PCR Based Diversity Studies

Effect of DNA extraction methods on the detection of porcine ingredients in halal cosmetics using real-time PCR

Diagnostic methods for detecting fungal pathogens on vegetable seeds

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