Research On DNA Extracting From The Faeces Of Wild Tarim Red Deer And Gender Identification

Tarim red deer (cervus elaphus yarkandensis) is listed as second category protected wildlife in china. It is unique among deer subspecies for adapting to desert environment, which has a very impotant status among XingJiang bio-diversity.It is only distributed in the narrow ranges in middle and lower reaches of Tarim River and lower reaches of Charchen River, Due to environmental degredation, the population of this subspecies have been urgently declined.Study on extracting DNA from faecal sample of wildlife animals,has attracted more attention at home and abroad in recent years.However because the faecal samples has so many characteristics.For example,having too many impuritie,seasily degraded,differences among individuals and so on. That often lead to the failure of DNA extraction.In this experiment, we established a faecal DNA extraction method that is suitable for Tarim red deer and larger herbivorous mammals by using non-invasive sampling method and based on combination of three types of domestic and foreign common characteristic of faecal DNA extraction method and discussed some relevant factors about faecal extraction. SRY and micro-satellite primers was used for PCR amplification of faecal DNA.To test the quantity of DNA and identification of the sex ration of wild population of the Tarim red deer.This experiment obtains the following results :(1)Samples were preserved by drying and freezing,and make multiple parallel tubes for each sample;faecal sample were preprocessed with actone or PBS ,and scalpel scraping were made for the external surface of faecal;CTAB was used for lysising the faecal cells,and SiO2 for specific adsorption of DNA.multiple tube products were merged and purified using purification kit at the last to obtain high quality DNA.(2)45 samples were successfully amplified from 50 faecal samples,among them 16 were male,29were famle,and the proportion of male and famle was 0.55:1.