Técnica no convencional de extracción de ADN a partir de tejido embebido en parafina para uso en la reacción en cadena de la polimerasa
Meza, Giovanna | 2013-01
The paraffin-embedded tissues are samples resting in the pathology banks, storing important genetic information, useful for retrospective studies of epidemiology and molecular pathology and diagnosis of diseases in humans and animals. Among the methods described for extracting DNA from tissues in paraffin, few are successful in obtaining the genetic material which can be amplified above 200bp fragments by the PCR technique. This paper describes the experience of U.D.C.A ́s molecular biology laboratory, related to the application of a methodology recently used to extract DNA from paraffin-embedded tissue and which permits to be optimal amplified by PCR of a fragment to 600pb. Two techniques were used, extensively referenced to these samples, such as the phenol extraction protocol and the commercial kit from QIAGEN®, and also a third commonly used method for extracting DNA from blood samples, but not for tissue in paraffin, the technique of extraction or salting out salts. The extracted DNA amplifiable ability was evidenced to obtain a fragment of 600pb establishing a human gene. The results showed that extraction with phenol and commercial kit from QIAGEN ® achieved amplifications of a 21%, while the salt extraction method a 91%. This work reveals that an unconventional technique of DNA extraction from paraffin-embedded tissue can offer better performance than traditional methodologies.