Rapid isolation of DNA from fossil and museum specimens suitable for PCR.
We describe a simple process for extraction of DNA from amber-entombed fossils and museum specimens that is suitable for enzymatic amplification by PCR. Five to ten milligrams of the macerated specimen were mixed in 300 microliters of silica matrix and shaken at 55 degrees C for 1 h in a sterile, screw-capped microcentrifuge tube. After incubation, the silica matrix was transferred to the upper chamber of a SpinFilter, centrifuged at maximum speed for 1 min and then washed twice with 500 microliters of wash solution and the DNA eluted with 50 microliters of TE buffer. The eluate was used as template for PCR, and the results were evaluated by electrophoresis and nucleotide sequence analysis. All samples tested yielded positive results, which were subsequently verified by sequence analysis. It appears, at least in our hands, that the procedure described here is a rapid and efficient way of obtaining small amounts of DNA for PCR in museum and fossilized specimens.
Cano
R J
RJ
Biological Sciences Department, California Polytechnic State University, San Luis Obispo 93407.
Poinar
H N
HN
D000818:Animals; D001483:Base Sequence; D001516:Bees / Q000235:genetics; D002498:Centrifugation; D004247:DNA / Q000737:chemistry / Q000302:isolation & purification*; D017931:DNA Primers; D004587:Electrophoresis, Agar Gel; D005580:Fossils; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA
D017931:DNA Primers; D004247:DNA