Extraction and amplification of DNA from formalin-fixed, paraffin-embedded tissues.
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.
Wu
Lin
L
Department of Human Genetics, Roche Molecular Systems, Inc., Alameda, California 94501, USA. Lin.Wu@roche.com
Patten
Nancy
N
Yamashiro
Carl T
CT
Chui
Buena
B
D001483:Base Sequence; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification*; D017931:DNA Primers / Q000235:genetics; D004273:DNA, Neoplasm / Q000235:genetics / Q000302:isolation & purification; D005260:Female; D005557:Formaldehyde; D016158:Genes, p53; D006358:Hot Temperature; D006801:Humans; D008297:Male; D009369:Neoplasms / Q000235:genetics; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction / Q000379:methods; D016707:Tissue Fixation
D017931:DNA Primers; D004273:DNA, Neoplasm; D005557:Formaldehyde; D004247:DNA