Publication: DNA extraction from paraff in-em bedded tissues using a salting-out procedure, a reliable method for PCR amplification of archiva1 material
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Date
1997
Authors
Howe, J.R. ; Klimstra, D.S. ; Cordon-Cardo, C.
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Publisher
Murcia : F. Hernández
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DOI
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info:eu-repo/semantics/article
Description
Abstract
Many techniques have been described for the
extraction of DNA from paraffin-embedded tissues.
Numerous efforts have been directed at simplification of
these methods for rapid analysis using PCR. One
disadvantage to some of the simpler procedures is
inefficient PCR amplification, and for more involved
ones using phenol/chloroform extraction, reduction in
the yield of DNA. In the present study we report the use
of a novel salting-out procedure that was utilized to
extract DNA from 259 separate microdissection
specimens of formalin-fixed, paraffin-embedded tissue
sections. These sections were derived from 97 patients
with tumors of the ampulla of Vater resected between
1965 and 1995 at our institution. The mean DNA yield
was 22.75 pg (median 13.2130.25) and the mean
2601280 absorbance ratio was 1.68 (median 1.7020.25).
Al1 specimens (2591259) were successfully used to
amplify K-ras exon 1 by a nested PCR technique. These
results indicate that this DNA extraction method
produces good yields of quality DNA, even from
specimens several decades old.
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