A simple and efficient method for extracting DNA from old and burned bone.
It has been a challenge to extract DNA from bones previously soaked in water, burned, or buried for a long time, due to the reduced quality and quantity of DNA in the bone samples. The dramatic degradation of the DNA and the presence of PCR inhibitors in the collagen significantly complicate the process of DNA identification in dated and charred bones. In this article, we present a novel strategy to obtain DNA from bones based on the use of cetyltrimethylammonium bromide (CTAB) lysis buffer and isoamyl alcohol-chloroform extraction with subsequent DNA purification using the DNA IQ System, or alternatively the QIAquick system. When applied to bones soaked, burned or buried for up to nine years, this method increases the purity and yield of DNA with respect to the traditional phenol-chloroform method and significantly improves multiplex STR genotyping using fluorescence-based methods. The results of this research will assist forensic scientists in the identification of DNA from victims whose bodies underwent significant trauma or burning, precluding the utilization of traditional forensic DNA identification techniques.
Ye
Jian
J
Institute of Forensic Science, Ministry of Public Security, Beijing, China. yejian_21@sina.com.cn
Ji
Anquan
A
Parra
Esteban J
EJ
Zheng
Xiufen
X
Jiang
Chengtao
C
Zhao
Xingchun
X
Hu
Lan
L
Tu
Zheng
Z
D001842:Bone and Bones / Q000737:chemistry*; D002021:Buffers; D000077286:Cetrimonium; D002593:Cetrimonium Compounds; D002725:Chloroform; D004247:DNA / Q000302:isolation & purification*; D016172:DNA Fingerprinting / Q000379:methods*; D018732:Forensic Anthropology / Q000379:methods*; D005838:Genotype; D006801:Humans; D000439:Pentanols; D012997:Solvents; D013501:Surface-Active Agents; D020080:Tandem Repeat Sequences
D002021:Buffers; D002593:Cetrimonium Compounds; D000439:Pentanols; D012997:Solvents; D013501:Surface-Active Agents; D002725:Chloroform; D004247:DNA; C029683:isopentyl alcohol; D000077286:Cetrimonium