| The species resources of China Macaca Mulatta had been very scarce and a reduction in the trend of investigation .In the field of law enforcement work and protection of the monkeys was often the case of suspected sample contamination, corruption, incomplete (such as hair, bone, hoof A, organ, muscle fibers, etc.) or a very small amount of samples, it was difficult to check the identification. It was very meaningful for the establishment of identified method to China Macaca Mulatta in molecular biology and technology,which in order to more effectively protect the monkeys of wild animals, to combat smuggling and illegal hunting.Test 1: In this study, the method used mainly non-invasive, extracted DNA from four species of monkeys fecal samples, downloaded cyt b gene sequences from Genbank of some common animals, five kinds of Macaca Mulatta and the similar to the case of the same family with head Branch animals, the use of software design and synthesis of monkeys was one pairs of universal primers (Ms / Ma), Macaca nemestrina primer 1 (Ts / Ta), Macaca mulatta primers (Ys /Ya), Macaca arctiodes primer 2 (Ds1/Da1, Ds2/Da2), Macaca assamensis primer 2 (Xs1/Xa1, Xs2/Xa2), and a large number of tests to their specificity, stability, and sensitivity.The results showed that: universal primer Ma/Ms is a species-specific, only the specific species of monkeys could amplify DNA, but some common animals and animals near the source of DNA can not be amplified; the other alsohad good specificity and stability. In particular, the sensitivity of tests to detect the minimum requirements of the species concentration.Test 2: Used Primer Express software,we designed and synthesized Taqman probe: Macaca nemestrina (Ts / Ta), Macaca mulatta 3 (Ms0/Ma0, Ms1/Ma1, Ms2/Ma2), Macaca arctiodes 1 (Ds1/Da1), Macaca assamensis (Xs1/Xa1), by a large number of tests, results showed that: Macaca mulatta probe (Ms0/Ma0), Macaca nemestrina probe (Ts / Ta) and Macaca arctiodes (Ds1/Da1) probe had good specificity, it could be separated with other Macaca mulatta. Monkeys probe (Ms1/Ma1, Ms2/Ma2) could first distinguish among Macaca mulatta, Macaca arctiodes and Macaca nemestrina, Macaca assamensis, and then used the probe (Ds1/Da1) could be distincted between Macaca mulatta and macaques. This test was first introduced a step-by-step method to identification, which be learned by the identification to other wild animals.Test 3: Extracted DNA from other samples: hair, muscle, blood, and fecal. Compared the consistency of the test results, which DNA template from different samples.The results showed:DNA from the hair, muscle, blood and fecal samples achieved the same result. It said that the method was stable and reliable, which aimed to the purpose of identification to microorganization of the monkeys.Test 4: run on agarose gel electrophoresis, compared the bands obtained and plastic recycling sequencing to see whether the outcome was the needed. We received the same results between sequence obtained from the test and the sequence from web publishing, the purpose is to strip. At the same time, designed sequencing primers, measured four species of monkeys parts of Cytb sequences, for subsequent extraction of the scientific basis for further study.The test established methods to the molecular biology of animal identification and laboratory confirmation of four kinds of Macaca mulatta. It was providing a convenient, sensitive and reliable means to detection when encountered animals and parts of trace residues of the inspection sample in field investigations.
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