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Related Experiment Videos

PCR-based apolipoprotein E genotype analysis from archival fixed brain

L Gioia1, L J Vogt, W M Freeman

  • 1Department of Physiology and Pharmacology, Wake Forest University School of Medicine, Winston-Salem, NC 27157-1083, USA.

Journal of Neuroscience Methods
|July 17, 1998
PubMed
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This study presents a new method for apolipoprotein E (apo E) genotyping from archived, formaldehyde-fixed tissues. The technique successfully determines apo E genotypes, aiding Alzheimer's disease research.

Area of Science:

  • Genetics
  • Neuroscience
  • Molecular Biology

Background:

  • Formaldehyde fixation of tissues can degrade DNA, hindering genetic analysis.
  • Apolipoprotein E (apo E) genotype is a significant risk factor for Alzheimer's disease (AD).
  • Retrospective analysis of archived samples is crucial for understanding AD pathogenesis.

Purpose of the Study:

  • To develop a reliable method for determining apolipoprotein E (apo E) genotype from archived, formaldehyde-fixed human tissues.
  • To enable retrospective studies on the association between apo E alleles and Alzheimer's disease.
  • To establish a robust DNA analysis protocol adaptable for various archived sample types.

Main Methods:

  • DNA extraction and purification from formaldehyde-fixed and archived tissues.

Related Experiment Videos

  • Nested polymerase chain reaction (PCR) amplification of the apolipoprotein E gene polymorphic region.
  • Genotyping using HhaI restriction fragment length polymorphism analysis of the amplified DNA.

    • Main Results:

    • Successful genotyping of 90% (34/38) of archived Alzheimer's disease patient brain samples.
    • The method was effective even for samples stored for up to 12 years in formalin.
    • The protocol demonstrated robustness with DNA from whole blood and frozen tissues.

    Conclusions:

    • The described nested PCR and RFLP method enables reliable apo E genotyping from challenging archived formalin-fixed tissues.
    • This technique facilitates retrospective genetic analysis, crucial for advancing Alzheimer's disease research.
    • The adaptable protocol can be applied to genotype other genes in various preserved biological samples.