NCJ Number
226990
Journal
Forensic Science International: Genetics Volume: 3 Issue: 2 Dated: March 2009 Pages: 67-73
Date Published
March 2009
Length
7 pages
Annotation
This paper reports on the design of five combinations of primer pairs for the identification of shells and shell fragments that may come from endangered and protected species of turtles that are illegally used as ornaments, food products, and medicines.
Abstract
A total of 58 samples of the 100 examined were amplified successfully using 2 primer pairs, L14724/H15197 and L14735/H15149, to amplify a partial section of the cyt b gene. All of the sequences reported were confirmed by repeated PCR amplifications and DNA sequencing using the forward and reverse primers. Of these 58 samples, 56 samples resulted in a sequence similarity greater than 98.0 percent when compared to a DNA sequence for a turtle species registered on GenBank/EMBL. The success rate for species identification using the test described is dependent upon the choice of primer sets used and the length of the expected amplification product. Gelled products were simulated by the process of decoction for up to 12 hours, after which all the turtle species could be identified from the liquid samples. The 100 turtle shells were identified previously as from 15 species based upon their morphology. They were provided by the Council of Agriculture in Taiwan and importers of Chinese herbal medicine. All 100 samples were submitted for DNA analysis using the cyt b gene, and their resulting DNA sequences were used in the design of universal primers for the amplification of smaller DNA fragments. This report describes the procedures of DNA extraction, PCR amplification and DNA sequencing, and simulation testing. 4 tables, 2 figures, and 41 references