Simple and Efficient Method for Extracting DNA From Old and Burned Bone

• NCJ Number: 206532
• Journal: Journal of Forensic Sciences Volume: 49 Issue: 4 Dated: July 2004 Pages: 754-759
• Author(s): Jian Ye B.S.; Anquan Ji M.S.; Esteban J. Parra Ph.D.; Xiufen Zheng Ph.D.; Chengtao Jiang B.S.; Xingchun Zhao B.S.; Lan Hu Ph.D.; Zheng Tu B.S.
• Date Published: July 2004
• Length: 6 pages
• Publication Format: Article
• Publication Type: Report (Study/Research)
• Language: English
• Country: United States of America

Annotation

This paper describes a new DNA isolation method for extracting DNA from old and burned bone by combining the use of cetyltrimethylammonium bromide (CTAB) buffer and isoamyl alcohol-chloroform, along with a further DNA purification step that uses the DNA IQtm System, or alternatively the QIAquicktm system.

Abstract

PCR amplification results are significantly affected by naturally occurring impurities in aged or burned bone samples. Moreover, DNA in aged or burned bone samples is significantly degraded, typically resulting in poor sample quantity and quality for STR examination. When extracting DNA from old bones, the major challenges are to maximize DNA yield, to obtain DNA of high quality, to eliminate PCR inhibitors, and to minimize the possibility of outside contaminants. The new method for extracting DNA from bone was tested with bone samples collected 2-9 years after death. Soaked and burned bones were also genotyped. All the samples were casework evidence analyzed at the Institute of Forensic Science in Beijing, China. Bone samples were surface sterilized by washing with 0.5 percent sodium hypochlorite and then rinsed with running deionized distilled water for 5 minutes. The bone samples were then air-dried and exposed to UV irradiation for 1 hour. Drilling into the material with a hand drill produced 1 to 3 g of bone powder, depending on the age and state of preservation. The experiment compared the efficiency of DNA extraction from bone using a traditional pheno-chloroform method and the novel CTAB plus isoamyl alcohol-chloroform method. Results indicated that the new method increased the purity and yield of DNA compared to the traditional method, and it significantly improved multiplex STR genotyping using fluorescence-based methods. Thus, this test of the new method proved it to be a stable, reliable, robust, and efficient strategy for DNA extraction from aged, soaked, and burned bones. 8 figures and 12 references