title: Automated DNA extraction from pollen in honey.
abstract: In recent years, honey has become subject of DNA analysis due to potential risks evoked by microorganisms, allergens or genetically modified organisms. However, so far, only a few DNA extraction procedures are available, mostly time-consuming and laborious. Therefore, we developed an automated DNA extraction method from pollen in honey based on a CTAB buffer-based DNA extraction using the Maxwell 16 instrument and the Maxwell 16 FFS Nucleic Acid Extraction System, Custom-Kit. We altered several components and extraction parameters and compared the optimised method with a manual CTAB buffer-based DNA isolation method. The automated DNA extraction was faster and resulted in higher DNA yield and sufficient DNA purity. Real-time PCR results obtained after automated DNA extraction are comparable to results after manual DNA extraction. No PCR inhibition was observed. The applicability of this method was further successfully confirmed by analysis of different routine honey samples.
authors.affiliation: Bavarian Health and Food Safety Authority, Molecular Biology, Veterinärstr. 2, 85764 Oberschleißheim, Germany. Electronic address: patrick.guertler@lgl.bayern.de.
authors.forename: Patrick
authors.initials: P
authors.lastname: Guertler
authors.forename: Adelina
authors.initials: A
authors.lastname: Eicheldinger
authors.forename: Paul
authors.initials: P
authors.lastname: Muschler
authors.forename: Ottmar
authors.initials: O
authors.lastname: Goerlich
authors.forename: Ulrich
authors.initials: U
authors.lastname: Busch
mesh_terms: D001331:Automation / Q000379:methods*; D005591:Chemical Fractionation / Q000379:methods*; D004247:DNA / Q000235:genetics / Q000302:isolation & purification*; D005506:Food Contamination / Q000032:analysis*; D006722:Honey / Q000032:analysis* / Q000382:microbiology; D011058:Pollen / Q000737:chemistry* / Q000382:microbiology; D016133:Polymerase Chain Reaction
chemical_list: D004247:DNA
keywords: Automated DNA extraction; CTAB; Honey; Maxwell; Pollen; Real-time PCR