A METHOD FOR THE EXTRACTION OF GENOMIC DNA FROM HUMAN BRAIN-TISSUE FIXED AND STORED IN FORMALIN FOR MANY YEARS

Citation
A. Savioz et al., A METHOD FOR THE EXTRACTION OF GENOMIC DNA FROM HUMAN BRAIN-TISSUE FIXED AND STORED IN FORMALIN FOR MANY YEARS, Acta Neuropathologica, 93(4), 1997, pp. 408-413
Citations number
10
Categorie Soggetti
Neurosciences,"Clinical Neurology",Pathology
Journal title
ISSN journal
00016322
Volume
93
Issue
4
Year of publication
1997
Pages
408 - 413
Database
ISI
SICI code
0001-6322(1997)93:4<408:AMFTEO>2.0.ZU;2-Z
Abstract
We report a method providing access to high molecular weight, polymera se chain reaction (PCR)-amplifiable genomic DNA from brains scored in formalin for many years. It consists mainly of an intensive proteinase K treatment of ground tissue previously embedded in agarose plugs, fo llowed by a washing and an elution step. The method was tested on brai ns fixed and stored in formalin for up to 46 years. All extracted DNA show an identical pattern of degradation ranging from well-preserved ( more than 20 kb) to 400-bp-long fragments. This was demonstrated for D NA extracted from the cerebellums of elderly psychiatric and geriatric patients (of more than 60 years of age), male and female, demented or not, with postmortem delays longer than 1 h and shorter than 1 day. I n all these cases PCR amplification of a 838-bp-long beta-actin produc t was successfully performed when proteinase K treatment was sufficien tly effective to generate pure DNA. Thus, high molecular weight, PCR-a mplifiable genomic DNA can be extracted from brains stored in formalin for almost half a century.