Simple and highly effective DNA extraction methods from old skeletal remains using silica columns
Received 1 June 2009; received in revised form 9 October 2009; accepted 10 October 2009. published online 20 January 2010. Corrected Proof
Abstract
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.
aDepartment 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
bHuman Identification Research Center, Yonsei University, 250 Seongsanno, Seodaemun-Gu, Seoul 120-752, Republic of Korea
Corresponding author at: Department of Forensic Medicine, Yonsei University College of Medicine, 250 Seongsanno, Seodaemun-Gu, Seoul 120-752, Republic of Korea. Tel.: +82 2 2228 2481; fax: +82 2 362 0860.