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Jornal Brasileiro de Patologia e Medicina Laboratorial
On-line version ISSN 1678-4774
NASCIMENTO, Ericson Martins; SPINELLI, Marilda Osti; RODRIGUES, Consuelo Junqueira and BOZZINI, Nilo. DNA extraction from paraffin material protocol in order to analyse microssatellites in uterine leiomyoma. J. Bras. Patol. Med. Lab. [online]. 2003, vol.39, n.3, pp. 253-255. ISSN 1678-4774. http://dx.doi.org/10.1590/S1676-24442003000300013.
The standardization of the protocol of the DNA extraction from paraffin material in order to analyze the instability of microsatellites in genes related to the uterine leiomyoma, using 30 to 40mg of tissue removed from paraffin blocks is presented. Desparaffin throught xilol baths to 65ºC, rehydrated with solutions of decreasing concentrations of ETOH and deionisation water. The extraction of DNA was obtained through the stages of cellular lysis, with Promega solution, digestion of proteins by Proteinase K, precipitation proteins with Promega solution and precipitation of DNA with ETOH 70% colded and ressuspended with Promega solution. This protocol results in a viable DNA to be use in the Polimerase Chain Reaction (PCR).
Keywords : Uterine leiomyoma; Instability of microsatellites; Paraffin material.