J Clin Pathol. 1990 Sep;43(9):748-51. Unique Identifier : AIDSLINE
The mechanism of DNA degradation and its clinical applications were
examined. When purified lambda phage and extracted liver DNA were fixed
in phosphate buffered formaldehyde, the DNA did not degrade, but there
was incomplete digestion with endonuclease. Rat liver tissues were fixed
under various conditions and DNA extracted. Immediate fixation with
buffered formaldehyde at low temperature, or the addition of EDTA to
buffered formaldehyde blocked the DNA degradation. Analysis of pulsed
field gel electrophoresis also showed that DNA was degraded before
extraction. These results suggest that tissue nuclease has an important
role in DNA degradation in tissue. Furthermore, formaldehyde fixation at
low temperature, which may take time and which decreases slightly the
staining capacity, is useful for the extraction of intact DNA. For
clinical application, the detection of provirus was examined. Genomic
DNA was extracted from a necropsy sample of adult T cell leukaemia fixed
in formaldehyde; human T cell leukaemia virus type-I (HTLV-I) provirus
was successfully detected by Southern blotting.
Animal DNA/DRUG EFFECTS/*METABOLISM DNA, Viral/ANALYSIS Edetic Acid
*Fixatives *Formaldehyde Human HTLV-I/GENETICS Middle Age Rats
JOURNAL ARTICLE