title: DNA extraction from fresh-frozen and formalin-fixed, paraffin-embedded human brain tissue.
abstract: Both fresh-frozen and formalin-fixed, paraffin-embedded (FFPE) human brain tissues are invaluable resources for molecular genetic studies of central nervous system diseases, especially neurodegenerative disorders. To identify the optimal method for DNA extraction from human brain tissue, we compared methods on differently-processed tissues. Fragments of LRRK2 and MAPT (257 bp and 483 bp/245 bp) were amplified for evaluation. We found that for FFPE samples, the success rate of DNA extraction was greater when using a commercial kit than a laboratory-based method (successful DNA extraction from 76% versus 33% of samples). PCR amplicon size and storage period were key factors influencing the success rate of DNA extraction from FFPE samples. In the fresh-frozen samples, the DNA extraction success rate was 100% using either a commercial kit (QIAamp DNA Micro) or a laboratory-based method (sample boiling in 0.1 mol/L NaOH, followed by proteinase K digestion, and then DNA extraction using Chelex-100) regardless of PCR amplicon length or tissue storage time. Although the present results demonstrate that PCR-amplifiable genomic DNA can be extracted from both fresh-frozen and FFPE samples, fresh brain tissue is recommended for DNA extraction in future neuropathological studies.
authors.affiliation: Neuroscience Research Australia and the University of New South Wales, Sydney, 2031, Australia, wangjh6304@sina.com.
authors.forename: Jian-Hua
authors.initials: JH
authors.lastname: Wang
authors.forename: Amany
authors.initials: A
authors.lastname: Gouda-Vossos
authors.forename: Nicolas
authors.initials: N
authors.lastname: Dzamko
authors.forename: Glenda
authors.initials: G
authors.lastname: Halliday
authors.forename: Yue
authors.initials: Y
authors.lastname: Huang
mesh_terms: D001921:Brain; D004247:DNA / Q000302:isolation & purification*; D005404:Fixatives; D005557:Formaldehyde; D005629:Frozen Sections; D006801:Humans; D016612:Paraffin Embedding; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016707:Tissue Fixation
chemical_list: D005404:Fixatives; D005557:Formaldehyde; D004247:DNA