In-house and interlaboratory validation of a method for the extraction of DNA from pollen in honey

Waiblinger, Hans‐Ulrich; Ohmenhaeuser, Marc; Meissner, Stefanie; Schillinger, Miriam; Pietsch, Klaus; Goerlich, Ottmar; Mankertz, Joachim; Lieske, Kathrin; Broll, Hermann

doi:10.1007/s00003-012-0774-z

Cited by 18 publications

(13 citation statements)

References 6 publications

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“…2. The higher DNA yield of extracts obtained with Wizard method is also emphasised comparing with other reports, which produced much lower values from honey samples extracted with CTAB method (9.5e39 ng/mL) (Waiblinger et al, 2012). In what concerns the different applied pre-treatments, Fig.…”

Section: Extraction Methodsmentioning

confidence: 53%

“…The mixture was stored at À20 C until subsequent DNA extraction. -Pre-treatment c was performed based on a protocol proposed by Waiblinger et al (2012) with some modifications. Fifty grams of honey sample were distributed into four sterile 50 mL centrifugation tubes (12.5 g honey per tube), followed by the addition of 40 mL of ultrapure water to each tube, stirring and incubation at 40 C for 10 min.…”

Section: Sample Preparationmentioning

confidence: 99%

“…In previous studies, DNA extraction of pollen from honey samples has been reported both by using in-house extraction methods or commercial DNA extraction kits. Cheng et al (2007) and Waiblinger et al (2012) both used CTAB-based extraction methods in order to isolate DNA from honey and evaluate the presence of genetically modified organisms. The use of DNeasy Blood and Tissue Kit (Qiagen GmbH) was reported in other works aiming at detecting DNA from different plant species in honey samples (Laube et al, 2010;Valentini, Miquel, & Taberlet, 2010).…”

Section: Introductionmentioning

confidence: 99%

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Improving DNA isolation from honey for the botanical origin identification

et al. 2015

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a b s t r a c tHoney is a natural product highly consumed due its known association with health benefits. Monofloral honeys are perceived as better quality products, being the most appreciated by consumers, thus attaining higher market values. Therefore efficient tools are needed as alternatives to the classical microscopic analysis presently used for the botanical origin identification of honey. In the present work, the use of DNA-based methods for the botanical species identification of honey is proposed. For this purpose, five DNA extraction methods (the kits NucleoSpin Plant (methods A and B) and DNeasy Plant Mini Kit, and the in-house CTAB-based and Wizard methods) combined with three different sample pre-treatments were applied to four honey samples (3 monofloral honeys of Calluna vulgaris, Lavandula spp. and Eucalyptus spp. and one multifloral honey). The 15 DNA extraction protocols were compared in terms of DNA integrity, yield and purity, as well as capacity of amplification targeting universal and adh1 specific genes of C. vulgaris. The results demonstrated the superior efficacy of the Wizard method in terms of DNA quality and amplification capacity, when combined with the sample preparation treatment with a mechanical disruption step of pollen to improve DNA yield. Although with considerable lower DNA yields, the CTAB and DNeasy methods were also successful because both were able to clearly amplify heather DNA from the monofloral heather honey.

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Section: Extraction Methodsmentioning

confidence: 53%

Section: Sample Preparationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Improving DNA isolation from honey for the botanical origin identification

et al. 2015

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“…Isolasi DNA metagenomik dilakukan dengan tiga metode. Pertama, isolasi dengan lisis sel secara secara langsung dari metode isolasi DNA metagenomik berdasarkan metode Kuske et al (1997) [7] [8] dan dipadukan dengan menggunakan PowerSoil DNA Isolation Kit. Sebanyak 1,2 gram sampel madu didistribusikan ke empat tabung Eppendorf 1,5 mL (0,3 gram madu per tabung), diikuti dengan penambahan 960 L air steril untuk setiap tabung lalu diinkubasi pada suhu 40 o C selama 10 menit.…”

Section: Isolasi Dna Metagenomikunclassified

AMPLIFIKASI FRAGMEN GEN 18S rRNA PADA DNA METAGENOMIK MADU DENGAN TEKNIK PCR (POLYMERASE CHAIN REACTION)

Prakoso¹,

Wirajana²,

Suarsa³

2017

IJLFS

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The aim of this research was to amplificate 18S rRNA gene fragment from honey’s metagenomic DNA using Polymerase Chain Reaction (PCR). The honey sample was collected from Seraya Tengah village, Karangasem regency. The best result of primer design from in silico test was continued to in vitro test using PCR method. The optimum conditions for amplification was obtained as follows: pre-denaturation at 95oC for 3 minutes and continued with 30 of amplification cycle (denaturation at 95°C for 1 minutes, annealing at 55°C for 1 minutes and elongation at 72°C for 1 minutes) and the last step continued with extension process at 72°C for 2 minutes. The size of DNA fragment band of amplified product was about 100 bp which obtained from the honey’s metagenomic DNA.

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“…With a collaboration of 14 labs in and around Germany has accompanied for the establishment of method. Finally, the in-house and interlaboratory validation evidenced the DNA extraction from five different samples (Honeydew Honey with multifloral honey, Wild flower honey "flowers of the mountain", Wild flower honey, Rape honey, Acacia-with multifloral honey) [38]. With the comparison of previous studies, an improved method for the efficient DNA extraction from honey samples were performed using commercial DNA isolation kits, finally stating that Wizard method with pretreatment has the maximum yield in the aspect of both purity and quantity [39].…”

mentioning

confidence: 99%

DNA as a Biomaterial in Diagnosis of Food Adulteration and Food Safety Assurance

Muthu¹

2017

RDMS

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Apart from general applications of DNA, it can also be used as tool to assure the food safety, which is the main aspect of this review. Increasing GMO productions also increased the concern regarding the adulteration of GMO food products in our daily life. Countries around the world, raising the regulations regarding the GMO products to the consumer's table. Subsequently, the mode of adulteration in food products with GMO products feels difficult to identify. The food products also adulterer with low cost products which can also be detected by the isolation and identification of DNA from that product. The main focus on this review is to discuss the utilization of DNA as an effective tool to detect the adulteration and GMO content in various food products like oil, honey, beverages, baked products, animal feed, fried products, adulteration of milk and milk products. In the field of food safety, microbial contamination is also a major problem. Isolation of DNA from food can also reveals the presence of microbial contamination in the earlier stage. Also, the review emphases the way of utilizing DNA in differentiating organic foods from non-organic foods. ISSN: 2576-8840How to cite this article: Duraimurugan K, Narendhran S, Manikandan M. DNA as a Biomaterial A food should be free from microbial contamination until it reaches the consumers table. All kind of safety measures must be ensured by industries to make it possible. But still microbes are found in food products by any source, leading to the spoilage of food product and causing food borne infections to the consumers. Home cooked food samples collected from the villages in Malawi has shown to be contaminated upto 35% of pathogenic microbes and by further investigation, it was found that contamination of food samples by poor handling and unhygienic conditions [11]. Microbial contamination is a crucial problem in various food industries causing major economic losses. Various industries like dairy, poultry industries follow various cutting-edge technologies to reduce the microbial contamination in end product [12,13]. Microbial contamination can be explored by various physical methods including microscopic and culture depend approaches. But these techniques have various limitations in the confirmation of microbial presence. In present decades, PCR based methods are highly adopted due to its high accuracy and reliability [14].In the present review, we discuss the utilization of DNA as an effective tool in ensuring the quality and safety in various foods and processed food products (Figure 1). DiscussionDifferent DNA isolation methods from food products [20]. Apart from the standard protocols and commercial kits, various modified protocols are also adopted by researchers in order to enhance the efficiency of quality and quantity of extracted DNA [21]. DNA based detection of adulteration and GMO in food productsMilk and milk products: Milk stands as a source of high protein based food for vegetarians around the world. Commercial milk is available in the f...

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In-house and interlaboratory validation of a method for the extraction of DNA from pollen in honey

Cited by 18 publications

References 6 publications

Improving DNA isolation from honey for the botanical origin identification

Improving DNA isolation from honey for the botanical origin identification

AMPLIFIKASI FRAGMEN GEN 18S rRNA PADA DNA METAGENOMIK MADU DENGAN TEKNIK PCR (POLYMERASE CHAIN REACTION)

DNA as a Biomaterial in Diagnosis of Food Adulteration and Food Safety Assurance

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