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�p���v��We proposed to develop a DNA testing method to distinguish cashmere fibers from those of yak and sheep, aiming to make a new item proposal to ISO/TC38/WG22. The cashmere products contaminating sheep and yak fibers have been sometimes displayed incorrectly. Thus, it is necessary to develop a method to efficiently differentiate fibers from cashmere, sheep, and yak. Because these three animals are different in species, difference(s) of nucleic acid sequence of genomic DNA can be reliably applied for the differentiation. Cashmere nucleotide sequences of DNA fragments of approximately 2 kb in length have been determined and were compared with those of sheep and yak. Approximately 1,000 or more candidates for the differentiation procedure were identified among the three animals. We also have found that some nucleotide alterations could be polymorphic within the same species, indicating they could not be used for the differentiation.
�@By testing several samples of cashmere products as a trial, dark black stain fiber sample was found to be difficult to be differentiated. Modification of reaction components upon proteolytic processing and using another enzyme for PCR were found to improve efficiency of the differentiation.
Proteomic analysis of the animal fibers was performed for verification and falsification for the DNA testing. We examined sample preparation of proteins from yak, cashmere and wool fibers. After the fibers were crushed and solubilized, protein molecules were separated by electrophoresis and were fragmented by the proteolytic enzyme treatment. Analysis of the resulted peptide fragments by TOF-MS revealed several peaks specific to yak, wool, or cashmere. By repeating the sample preparation procedures, reproducible identification of the specific peaks were confirmed. We also improved some procedures for crushing fiber samples.
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