By Peter Bedson, Helen Parkes, V (Ginny) Saunders, Sue Upton
Introduces the problems of validation and caliber to the bioanalytical group, particularly addressing DNA-based analyses. Designed to elevate information of the standards which can impression the validity of DNA research and the construction of extra demanding situations which are linked to the research of actual samples. Spiral.
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Additional info for Analytical Molecular Biology
A clear understanding of what is required of the analytical data is vital to applying the correct technical and methodological approach. Verify whether qualitative, semi-quantitative or quantitative end-point analysis is required. 4) required of a method. 2. Assess the sample. Confirm the nature of the sample (matrix, size) and form a strategy for sample collection, transportation, storage and preparation. 3). Ensure a working sample identification system is in place so that each sample has a unique identifier.
1, where DNA was extracted from meat samples. Equal amounts of four types of bovine tissue (A, steak; B, kidney; C stewing steak; D, fat) were extracted using a typical validated method. At the stage prior to DNA precipitation, the extract was divided into four and precipitated as described before an aliquot was electrophoresed on an agarose gel and visualised under UV light. 1 demonstrates how the alteration of a single procedure in the extraction process can alter the result obtained. In this instance, one of the four alcohol precipitations carried out (lanes 3) significantly reduced the yield of the D N A obtained.
Such precautions will prevent contamination and further degradation of the sample. Factors affecting the validity of DNA extraction due to sample storage have been identified as: D N A Extraction 0 0 35 Incorrect sample storage temperature - Further growth of contaminating microorganisms can be halted by freezing a sample. Naturally occurring autolysis and DNA degradation by endogenous and exogenous enzymes can also be abated by reduced temperatures. Samples requiring long-term storage should either be placed at - 80 "C, preferably after snap cooling (fast freezing in liquid nitrogen), or lyophilised,' after which a sample can be stored at room temperature in a dry environment for extended periods.