Analysis of DNA from degraded tissue

Abstract

To investigate the PCR-amplified fragments sizes generated from DNA templates obtained from degraded tissue samples at various stages, we simulated conditions leaving 2-inch cubic chunks of pork to decompose for 1 week in water, sea-water, and air-dried. Tissue samples were collected everyday and PCR analyzed using the nuclear β-actin gene and one from the mitochondrial cytochromeB gene primers. It was shown that the 211-, 289- and 366-bp nuclear β-actin DNA fragments were amplified from all samples immersed in water and sea water up to the 8th day. The 289-bp fragment was amplified from samples, which were left to air-dry up to the 4th day. The 323-bp mitochondrial cytochrome B fragment was amplified from all samples up to the 8th day and PCR-amplified from all air-dried samples when the experiment was extended up to the 14th day. The results suggested that there is a good chance to obtain a full nuclear DNA profile from tissue samples immersed in the water and sea water for a week. However, if tissue samples were left air-dried in open environment over 4 days, there would be a high chance to obtain an incomplete/ no DNA profile. © 2008 Elsevier Ireland Ltd. All rights reserved.