ABSTRACT
Objective: To establish a method of restriction fragment length polymorphism (RFLP) to determine the origin of sika deer bones.Methods: The DNA in the bone samples was extracted after decalcification, and then amplified using polymerase chain reaction (PCR).The origin of the samples was further identified using RFLP analysis.Results: The bone samples of sika deer and red deer could be distinguished from those of pig, bovine and dog by PCR.And the samples of sika deer and red deer could be further distinguished by RFLP through the analysis of the length of restriction enzyme XbaI.Conclusion: A RFLP method is established to determine the origin of sika deer bones.
ABSTRACT
<p><b>OBJECTIVE</b>To establish a convenient, practical and high efficient method of DNA extraction of os cervi, and lay the foundation of identification of animal bones.</p><p><b>METHOD</b>The bones of sika deer, red deer, cattle, dog and pig were used to extract DNA under different decalcification time (24,48,72 h) and decalcification temperature (4,25,37,56,70 degrees C), and extract method.</p><p><b>RESULT</b>It proved by experiments that demineralization process promotes the cracking of osteocyte. In a broad of decalcification time and temperature, DNA could be extracted from all bone samples successfully while the quantity varied slightly.</p><p><b>CONCLUSION</b>Samples (about 0.1 g) decalcify with 0. mol x L(-1) EDTA at 4 degrees C for 24 h, then water-bath for 1 h after lysis buffer added, DNA extracted via the method above is of high quality and can be used for PCR.</p>