Extraction, evaluation, and amplification of DNA from decalcified and undecalcified United States Civil War bone.
Deoxyribonucleic acid (DNA) was extracted from documented skeletal specimens of U.S. Civil War soldiers to determine the need for decalcification prior to extraction. The polymerase chain reaction (PCR) was performed to determine if the calcification state had an effect on the ability to amplify the extracts and to determine how successful amplification would be with these aged specimens. Bone samples were pulverized to a fine powder and divided into two sets. One set of samples was decalcified and the other set left undecalcified. Both sets were extracted using an organic procedure. The results demonstrate that decalcification is not a necessary step in the extraction process and that the yield of DNA is generally two times greater when decalcification is omitted. Furthermore, the calcification state had no effect on the ability to perform the PCR. Although the extracted DNA was very degraded, a 410 base pair (bp) segment of the mitochondrial DNA (mtDNA) control region was amplified. These results suggest that DNA can be extracted and amplified from 125 year old bone without decalcification, which may assist in the identity of modern and historic forensic specimens.
Fisher
D L
DL
Armed Forces DNA Identification Laboratory, Armed Forces Institute of Pathology, Washington, DC.
Holland
M M
MM
Mitchell
L
L
Sledzik
P S
PS
Wilcox
A W
AW
Wadhams
M
M
Weedn
V W
VW
D000885:Anthropology, Physical / Q000379:methods*; D001483:Base Sequence; D001842:Bone and Bones / Q000737:chemistry*; D002113:Calcification, Physiologic; D004247:DNA / Q000032:analysis*; D004587:Electrophoresis, Agar Gel; D005784:Gene Amplification; D049672:History, 19th Century; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D014481:United States; D014857:Warfare
D004247:DNA