PCR-CTPP: a rapid and reliable genotyping technique based on ZFX/ZFY alleles for sex identification of tiger (Panthera tigris) and four other endangered felids
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An inexpensive, time-saving and reliable method, polymerase chain reaction with confronting two-pair primers (PCR-CTPP), was developed for sex identification in tiger (Panthera tigris) based on zinc finger alleles (ZFX/ZFY). A site of “C/G” transversion representing fixed differences that discriminated between ZFX and ZFY exons among felids was identified for primers designing. This primer set was successfully tested on samples including blood, shed hairs, dried skin, and stool which contained potential contamination caused by prey DNA. Cross species tests shown that this primer set was also useful for sex identification in four other endangered felids.
KeywordsFelids PCR-CTPP Sex identification Tiger ZFX ZFY
We are grateful to Chongqing Zoo, Guangzhou Zoo, Fuzhou Zoo, Nanchang Zoo, Changsha Zoo, Luoyang Zoo, Shanghai Zoo, Suzhou Zoo, Xiamen Zoo, Jiujiang Zoo, Wuhan Zoo, Guiyang Zoo, Chengdu Zoo, Kunming Zoo, Urumchi Zoo, Lanzhou Zoo, Xining Zoo, Beijing Zoo, Guizhou Museum and Heilongjiang Siberia Tiger Park, for providing the samples for this study. We also thank Ou Li and Emily H. King from Sichuan University, and two anonymous reviewers for amendments and comments on this manuscript. For financial support we acknowledge the Research Fund for Giant Panda Breeding of Chengdu, North of England Zoological Society, and the Chinese Association of Zoological Gardens (CAZG).
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