Sample Preparation and DNA Extraction for the Detection of Allergenic Nut Materials
Hernan Valdivia and Malcolm Burns1, Molecular and Cell Biology, LGC, Queens Road, Teddington, Middlesex, TW11 0LY, UK.
1 To whom all correspondence should be sent. E-mail: Malcolm.Burns@lgc.co.uk , Telephone: (+44) (0)208 943 7000, Fax: (+44) (0)208 943 2767
DNA analysis can be thought of as a series of procedural steps, starting with, for example, sampling, sample preparation, DNA extraction, DNA quantitation, PCR setup, equipment operation, software analysis, manual analysis, and user interpretation. Measurement uncertainty can affect any of these procedural steps, but the upstream stages of sampling and sample preparation are known to have a large effect upon the confidence with which a result is expressed.
We describe here an analytical approach for the effective sample preparation of nuts, to maximise the quantity and quality of DNA extracted from raw nut materials. The approach includes the de-shelling of nut material, novel sample treatment and grinding, and subsequent DNA extraction. The “fitness for purpose” of this approach was verified by measuring the quantity and quality of the DNA using UV spectrophotometry and subsequent PCR tests.
Keywords: DNA, DNA extraction, nuts, allergens, sample preparation
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