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《Journal of Anhui Normal University(Natural Science)》 2008-02
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Optimization of Experiment Conditions and Primer Screening With SRAP Markers for Chinese Alligators

TIAN Ming-li,WU Xiao-bing(College of Life Sciences,Anhui Normal University,Wuhu 241000,China)  
Chinese alligator(Alligator sinensis)is a critically endangered endemic species under legislative protection.Selecting suitable SRAP-PCR primers and establishing the optimal reaction system is the base of Alligator wild populations and captive population genetic research.The concentrations of Mg2+,dNTPs,Taq DNA polymerase,primers which affect the SRAP-PCR reactions were optimized in order to establish the SRAP molecular marker system in Chinese Alligator.The effects of primer pairs,whole DNA concentration,Mg2+ concentration,Taq DNA polymerase concentration and dNTPs concentration on SRAP reaction system were studied.Altogether 13 primer pairs were selected which can amplify unequivocally visible,monomorphic bands.And we accessed a stable,repeatable SRAP-PCR reaction system and the best of PCR parameters.The results showed that in 30μl PCR reaction: template DNA 1μl(30ng/μl),Mg2+(2.5mmol/L)2μl,dNTPs(2.5mmol/L)2μl,each primer(10pmol/μl)1μl;Taq DNA polymerase 1 U,The work provides a standardized procedure and powerful tool for molecular biology research of the Chinese Alligator.
【Fund】: 国家自然科学基金(30470244 30270213);; 安徽省学术与技术带头人专项基金;; 安徽省重要生物资源保护与利用安徽省重点实验室基金
【CateGory Index】: Q78
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