Automated DNA preparation from maize tissues and food samples suitable for real-time PCR detection of native genes

Hahnen, Silke; Offermann, Sascha; Miedl, Brigitte; Rüger, Barbara; Peterhänsel, Christoph

doi:10.1007/s00217-002-0579-x

Cited by 5 publications

(3 citation statements)

References 8 publications

(7 reference statements)

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“…Despite having low DNA yield and displaying faint smears in the gel images but having good purity overall, the MagNA Pure method seem to have amplified well in the screening stage. This is consistent with another study (Hahnen, Offermann, Meidl, Ruger, & Peterhansel, 2002) that also used the MagNA Pure LC purification system for DNA extraction from maize tissue and food samples where no DNA was visible in the gel image but samples were successfully amplified for all targets. This suggested that automated DNA preparation with Isolation Kit I which is actually optimized for the isolation of genomic DNA from mammalian whole blood and cultured cells allows extraction of good quality DNA from maize as well as highly processed feeds.…”

Section: Dna Extractionsupporting

confidence: 91%

Real-time PCR-based detection and quantification of genetically modified maize in processed feeds commercialised in Malaysia

et al. 2010

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a b s t r a c tThe present study which dealt mainly with processed feeds and some maize samples sold commercially in Malaysia evaluated the implementation of a real-time PCR cycling system for singleplex screening of eight target sequences (lectin, hmg, adh1, p35S, NK603, GA21, MON810 and MON863) and quantification of four genetically modified (GM) maize events (NK603, GA21, MON810 and MON863). The effects of using proprietary glass magnetic particles to bind DNA to their surface were also investigated in terms of DNA quantity, purity, integrity, quality and its overall effect on DNA amplification. GM material was present in 26.2% feeds and 65% maize samples. All GM samples contained MON810 followed by NK603 (47.5%), GA21 (25%) and MON863 (2.5%). Single-event and multiple-events were identified in the GM samples with 50% of the GM samples containing multiple-events. The present study which represents a fast and reliable methodology would provide an overview of the presence and levels of GMOs in feeds and maize in Malaysia.

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Section: Dna Extractionsupporting

confidence: 91%

Real-time PCR-based detection and quantification of genetically modified maize in processed feeds commercialised in Malaysia

et al. 2010

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“…The ability to monitor the progress of the PCR in real time completely revolutionized the approach to PCRbased quantitation of DNA and RNA. Compared to other endpoint quantitation methods, real-time PCR offers a streamlined assay development, reproducible results, and a large dynamic range, and it has good specificity and sensitivity (22).…”

Section: Resultsmentioning

confidence: 99%

Qualitative and Quantitative Evaluation of the Genomic DNA Extracted from GMO and Non-GMO Foodstuffs with Four Different Extraction Methods

Peano

Samson

Palmieri

et al. 2004

J. Agric. Food Chem.

119

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The presence of DNA in foodstuffs derived from or containing genetically modified organisms (GMO) is the basic requirement for labeling of GMO foods in Council Directive 2001/18/CE (Off. J. Eur. Communities 2001, L1 06/2). In this work, four different methods for DNA extraction were evaluated and compared. To rank the different methods, the quality and quantity of DNA extracted from standards, containing known percentages of GMO material and from different food products, were considered. The food products analyzed derived from both soybean and maize and were chosen on the basis of the mechanical, technological, and chemical treatment they had been subjected to during processing. Degree of DNA degradation at various stages of food production was evaluated through the amplification of different DNA fragments belonging to the endogenous genes of both maize and soybean. Genomic DNA was extracted from Roundup Ready soybean and maize MON810 standard flours, according to four different methods, and quantified by real-time Polymerase Chain Reaction (PCR), with the aim of determining the influence of the extraction methods on the DNA quantification through real-time PCR.

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“…Binding and elution from silica has also become the procedure of choice for most nucleic acid extraction procedures (Smith et al 2003 ). In addition, many different commercial kits and automated procedures for food samples are currently available (Marmiroli et al 2008 ), although the automation is restricted nowadays to a limited number of food samples (Hahnen et al 2002 ). Several papers compare different extraction methods (Zimmermann et al 1998a ;Peano et al 2004 ).…”

Section: Dna -Based Methodologies For Gmo Analysismentioning

confidence: 99%

Assessment of Genetically Modified Organisms (GMO) in Meat Products by PCR

Hernández

Ferrando

Rodrı́guez-Lázaro

2010

Handbook of Meat Processing

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Automated DNA preparation from maize tissues and food samples suitable for real-time PCR detection of native genes

Cited by 5 publications

References 8 publications

Real-time PCR-based detection and quantification of genetically modified maize in processed feeds commercialised in Malaysia

Real-time PCR-based detection and quantification of genetically modified maize in processed feeds commercialised in Malaysia

Qualitative and Quantitative Evaluation of the Genomic DNA Extracted from GMO and Non-GMO Foodstuffs with Four Different Extraction Methods

Assessment of Genetically Modified Organisms (GMO) in Meat Products by PCR

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