Abstract

BAENA DEL VALLE, Javier Alonso; RAMOS MORENO, Álvaro José; GOMEZ ALEGRIA, Claudio Jaime  and  GOMEZ CAMARGO, Doris Esther. Comparison of DNA extraction methods from formalin-fixed paraffin sections for RCP amplification. Rev. colomb. biotecnol [online]. 2013, vol.15, n.1, pp.172-179. ISSN 0123-3475.

Formalin-fixed and paraffin-embedded tissues are a source of important molecular findings in clinical and scientific, demonstrating that the DNA extracted from these is suitable for amplification by polymerase chain reaction (PCR). In this study, we tested three methods of DNA extraction, in order to compare the efficiency of these DNA for RCP amplification. Three samples were used, corresponding to a lung biopsy, endometrial curettage and lymph node, all fixed in 10% formaldehyde and embedded in paraffin. Three different methods were used for DNA extraction (extraction by salting out, modified Sambrook method and commercial kit) The DNA obtained was analyzed by spectrophotometry, and gel electrophoresis was performed in 1% agarose to check if the DNA was amplifiable. PCR was performed for exon 3 of caveolin-1 gene. All methods resulted in a good product of genomic DNA, obtaining more quality and purity in the salting out and commercial kit methods. Also, we obtained amplification of the product by these two methods, without favorable results with the DNA extracted by the modified "Preparation of Genomic DNA from Mouse Tails and Other Small samples, according to Sambrook et al." The DNA obtained from FFPE can be amplified by several methods, among them, salting out extraction is an easy, effective and low toxicity for obtaining good quality DNA for PCR amplification.

Keywords : DNA; isolation & purification; paraffin embedding; formaldehyde; polymerase chain reaction.

        · abstract in Spanish     · text in Spanish     · Spanish ( pdf )