DNA extraction from formalin-fixed, paraffin-embedded tissues: protein digestion as a limiting step for retrieval of high-quality DNA
Description
Several DNA extraction methods have been used for formalin-fixed, paraffin-embedded tissues, with variable results being reported regarding the suitability of DNA obtained from such sources to serve as template in polymerase chain reaction (PCR)-based genetic analyses. We present a method routinely used for archival material in our laboratory that reliably yields DNA of sufficient quality for PCR studies. This method is based on extended proteinase K digestion (250 micrograms/ml in an EDTA-free calcium-containing buffer supplemented with mussel glycogen) followed by phenol-chloroform extraction. Agarose gel electrophoresis of both digestion buffer aliquots and PCR amplification of the beta-globin gene tested the suitability of the retrieved DNA for PCR amplification.
Links
- http://scholar.google.co.uk/citations?view_op=view_citation&hl=en&user=b632u8sAAAAJ&pagesize=100&sortby=pubdate&citation_for_view=b632u8sAAAAJ:u5HHmVD_uO8C
- https://www.researchgate.net/publication/13721713_DNA_extraction_from_formalin-fixed_paraffin-embedded_tissues_protein_digestion_as_a_limiting_step_for_retrieval_of_high-quality_DNA
- http://dx.doi.org/10.1097/00019606-199712000-00006
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Published on 16 Nov 2012 - 21:08 (GMT)
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Authors
- Salvador J. Diaz-Cano
- S P Brady
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