Abstract

TISCORNIA, María M et al. Standarization of a method for paraffin-embedded clinic samples DNA extraction. Rev. cienc. tecnol. [online]. 2010, n.14, pp.53-57. ISSN 1851-7587.

Molecular Epidemiology includes genotypic studies of different biological processes, including pathological diseases such as cancer. Therefore, the paraffin-embedded clinical samples analysis is very important for diagnosis, prevention and therapy. Previously, many problems associated with amplification of DNA obtained from paraffin-embedded tissue samples were reported. This paper compares the capability of nine methodologies for DNA extraction from paraffin-embedded clinical samples, amplifying it by PCR to analyze under-250-bp fragments corresponding to folate pathway enzymes polymorphisms. Modification of "salting-out" method with concentrations of 0.5 M NaCl and 2 % CTAB detergent and Proteinase K was the most suitable for amplifiable DNA obtention of selected amplicons. All methods showed better amplification with longer cycles and lower annealing temperatures as compared to less fragmented DNA samples.

Keywords : Paraffin-embedded samples; DNA extraction.

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