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Extraction and amplification of DNA from formalin-fixed, paraffin-embedded tissues
Lin Wu1, Nancy Patten, Carl T Yamashiro
1Department of Human Genetics, Roche Molecular Systems, Inc., Alameda, California 94501, USA. Lin.Wu@roche.com
Abstract:
Formalin-fixed, paraffin-embedded tissue (PET) is an invaluable resource for retrospective molecular genetic studies, but the extraction of high-quality genomic DNA from the PET may be problematic. We report a simple method that significantly improves the ability to amplify DNA recovered from formalin-fixed PET. Based on the standard procedure of a commercially available DNA preparation kit, the QIAamp DNA mini kit or the HighPure DNA preparation kit, we developed this method by eliminating the xylene/ethanol extraction step and adding a heat-treatment step. With this method, we have observed a five- to 10-fold increase in amplification efficiency of a fragment in a range of 90 to 386 base pairs. We also have obtained much higher amplification efficiencies for a multiplex polymerase chain reaction.