DNA extraction from paraffin-embedded tissues using a salting-out procedure: a reliable method for PCR amplification of archival material.
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 micrograms (median 13.2 +/- 30.25) and the mean 260/280 absorbance ratio was 1.68 (median 1.70 +/- 0.25). All specimens (259/259) 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.
Howe
J R
JR
Department of Surgery, Memorial Sloan-Kettering Cancer Center, New York, NY 10021, USA.
Klimstra
D S
DS
Cordon-Cardo
C
C
D014670:Ampulla of Vater / Q000737:chemistry; D001483:Base Sequence; D004247:DNA / Q000737:chemistry / Q000302:isolation & purification*; D011905:Genes, ras / Q000235:genetics; D006801:Humans; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction / Q000379:methods*; D017421:Sequence Analysis; D013995:Time
D004247:DNA