Improved method facilitates reliable APOE genotyping of genomic DNA extracted from formaldehyde-fixed pathology specimens.
Apolipoprotein E (APOE) genotyping of genomic DNA extracted from formaldehyde-fixed specimens is cumbersome: there is not only a low yield or failure of PCR amplification (presumably due to degradation of DNA in the formaldehyde-fixed and paraffin-embedded tissue), but the standard method also involves the separation of DNA fragments as small as 48, 72, 81 and 91 bp requiring high-yield PCR products. Here we report about a semi-nested PCR method suitable for providing specific high-yield PCR products from DNA that has been extracted from formaldehyde-fixed specimens which initially generate low-quality templates. This method facilitates reliable APOE genotyping of DNA from difficult templates.
Ghebremedhin
E
E
Department of Anatomy, J.W. Goethe-University, Frankfurt/Main, Germany.
Braak
H
H
Braak
E
E
Sahm
J
J
D001057:Apolipoproteins E / Q000032:analysis* / Q000235:genetics*; D001344:Autopsy; D004247:DNA / Q000032:analysis* / Q000235:genetics*; D005557:Formaldehyde; D016678:Genome; D005838:Genotype; D006801:Humans; D016133:Polymerase Chain Reaction; D016707:Tissue Fixation
D001057:Apolipoproteins E; D005557:Formaldehyde; D004247:DNA