DNA extraction by sonication: a comparison of fresh, frozen, and paraffin-embedded tissues extracted for use in polymerase chain reaction assays.
DNA extraction from fixed tissues can be the most laborious and complex step in amplifying DNA by the polymerase chain reaction (PCR). We have previously reported a rapid and efficient method for extracting DNA by the use of sonication and glass beads. We have extended our experiences with this technique using fresh, frozen, and formalin-fixed paraffin-embedded tissues with and without the use of glass beads and report their results. Multiple tissue types were obtained at autopsy or as part of a surgical specimen. DNA was extracted from identical tissue when the sample was fresh, frozen, or formalin-fixed paraffin-embedded. Our results indicate that in most instances the sonication technique, which takes only 30 min from start to finish, can rapidly extract fresh, frozen, or formalin-fixed paraffin-embedded tissue and is superior to other rapid extraction techniques in terms of quality and quantity of DNA. It is much more rapid than those techniques that use long digestion periods. This technique will be of great value to those investigators extracting DNA for polymerase chain reaction assays.
Heller
M J
MJ
Department of Pathology, University of Iowa College of Medicine, Iowa City.
Robinson
R A
RA
Burgart
L J
LJ
TenEyck
C J
CJ
Wilke
W W
WW
D004247:DNA / Q000032:analysis*; D005615:Freezing; D005898:Glass; D006801:Humans; D008863:Microspheres; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction; D013010:Sonication
D004247:DNA