An efficient method for the extraction of DNA from formalin-fixed, paraffin-embedded tissue by sonication.
A method was developed for fast and efficient isolation of DNA from formalin-fixed, paraffin-embedded tissue sections for subsequent use in PCRs and DNA hybridization assays. The method relies on the use of a sonicating water bath to disrupt tissue samples to which a small amount of micro-sized glass beads have been added. The sonicating glass beads provide fast and efficient physical shearing of fixed tissue sections, allowing for quick release and solubilization of the DNA. The extraction process from paraffin section to amplifiable target DNA takes 30 minutes. The method eliminates the need for repetitive solvent extractions and exhaustive proteinase K digestion. PCR amplification of human genomic and viral target sequences was successfully carried out on DNA isolated from a number of different types of normal and infected tissues.
Department of Pathology, University of Iowa, College of Medicine, Iowa City 52242.
M J
MJ
Heller
L J
LJ
Burgart
C J
CJ
TenEyck
M E
ME
Anderson
T C
TC
Greiner
R A
RA
Robinson
D001483:Base Sequence; D004247:DNA / Q000302:isolation & purification*; D004279:DNA, Viral / Q000302:isolation & purification; D005557:Formaldehyde; D005821:Genetic Techniques; D006801:Humans; D008969:Molecular Sequence Data; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction; D013010:Sonication; D013997:Time Factors
D004279:DNA, Viral; D005557:Formaldehyde; D004247:DNA