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Nested amplification of COL2A1 3' variable region in skeletal remains
K Honda1, E Sugiyama, A Tsuchikane
1Department of Legal Medicine, Shinshu University School of Medicine, Nagano, Japan.
Nihon Hoigaku Zasshi = the Japanese Journal of Legal Medicine
|June 1, 1994
Summary
This study introduces a highly sensitive nested PCR method for forensic identification. It effectively amplifies degraded DNA from skeletal remains, aiding in the identification of unidentifiable bodies.
Area of Science:
- Forensic Science
- Molecular Biology
- Genetics
Background:
- Skeletal remains are often highly decomposed and contaminated.
- DNA extraction from such samples presents significant challenges for identification.
Purpose of the Study:
- To develop a highly sensitive method for amplifying degraded DNA from skeletal remains.
- To improve the forensic identification of old, unidentifiable skeletal remains.
Main Methods:
- DNA was extracted from degraded skeletal remains (teeth and bone fragments).
- The COL2A1 3' variable region was amplified using a hypersensitive nested PCR technique.
- Nested PCR involved 47 cycles of amplification, significantly increasing sensitivity and specificity.
Main Results:
- The nested PCR method demonstrated 2,000 times greater sensitivity than standard 27-cycle PCR.
- The technique successfully amplified target DNA fragments from as little as 5 pg of template DNA.
- Agarose gel electrophoresis with ethidium bromide staining confirmed fragment detection.
Conclusions:
- Hypersensitive nested PCR is a suitable and effective method for the forensic identification of old, unidentifiable skeletal remains.
- This technique enhances the ability to obtain genetic profiles from challenging forensic samples.