Simple and highly effective DNA extraction methods from old skeletal remains using silica columns.
The recovery of DNA data from old skeletal remains is often difficult due to degraded and very low yield of extracted DNA and the presence of PCR inhibitors. Herein, we compared several silica-based DNA extraction methods from artificially degraded DNA, DNA with PCR inhibitors and DNA from old skeletal remains using quantitative real-time PCR. We present a modified large-scale silica-based extraction combined with complete demineralization, that enables maximum DNA recovery and efficient elimination of PCR inhibitors. This is performed with high concentration of EDTA solution for demineralization of bone powder followed by QIAamp spin columns and buffers from the QIAquick PCR purification kit. We have successfully used this modified technique to perform STR analysis for 55-year-old skeletal remains. The results of this study will contribute to solve the forensic cases dealing with skeletal remains.
Lee
Hwan Young
HY
Department of Forensic Medicine and Brain Korea 21 Project for Medical Science, Yonsei University College of Medicine, 250 Seongsanno, Seodaemun-Gu, Seoul 120-752, Republic of Korea.
Park
Myung Jin
MJ
Kim
Na Young
NY
Sim
Jeong Eun
JE
Yang
Woo Ick
WI
Shin
Kyoung-Jin
KJ
D001842:Bone and Bones / Q000378:metabolism*; D004247:DNA / Q000302:isolation & purification*; D006801:Humans; D018895:Microsatellite Repeats; D016133:Polymerase Chain Reaction / Q000379:methods
D004247:DNA