|本期目录/Table of Contents|

[1]莫然,谢仰杰,肖志群,等.中国鲎微卫星引物扩增圆尾鲎DNA的PCR反应体系优化[J].集美大学学报(自然科学版),2012,17(2):89-95.
　MO Ran,XIE Yang-jie,XIAO Zhi-qunWENG Zhao-hong.Optimization of a PCR Reaction System of Carcinoscorpius rotundicauda DNA Amplified by Microsatellite Primers from Tachypleus tridentatus[J].Journal of Jimei University,2012,17(2):89-95.
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《集美大学学报（自然科学版）》[ISSN:1007-7405/CN:35-1186/N]

卷:
第17卷
期数:
2012年第2期
页码:
89-95
栏目:
水产科学与生物工程
出版日期:
2012-03-25

文章信息/Info

Title:
Optimization of a PCR Reaction System of Carcinoscorpius rotundicauda DNA Amplified by Microsatellite Primers from Tachypleus tridentatus
作者:
莫然1; 谢仰杰1; 2; 肖志群1; 2; 翁朝红1; 2; 3
(1.集美大学水产学院,福建 厦门 361021;2.福建省高校水产科学技术与食品安全重点实验室,福建 厦门 361021;3.浙江海洋学院水产学院,浙江 舟山 316004)
Author(s):
MO Ran1; XIE Yang-jie12; XIAO Zhi-qun12WENG Zhao-hong1; 23
(1.Fisheries College,Jimei University,Xiamen 361021,China;2.Key Laboratory of Science and Technology for Aquaculture and Food Safety(Jimei University),Fujian Province,Xiamen 361021,China3.Fisheries College of Zhejing Ocean University,Zhoushan 316004,China)
关键词:
圆尾鲎; 中国鲎; 微卫星引物; PCR; 正交优化
Keywords:
Carcinoscorpius rotundicauda; Tachypleus tridentatus; microsatellite primers; PCR;  orthogonal optimization
分类号:
-
DOI:
-
文献标志码:
-
摘要:
        采用正交设计法，从dNTPs浓度、引物浓度、Mg2+浓度、Taq DNA聚合酶用量4个因素3个水平出发，优化设计圆尾鲎DNA的PCR反应体系（引物为中国鲎微卫星引物）.并采用直观分析方法分析正交试验结果，最终建立了圆尾鲎SSR-PCR最佳反应体系：总体积20 μL,Taq DNA聚合酶1.5 U、dNTPs 0.16 mmol/L、引物0.2 μmol/L、Mg2+2.0 mmol/L；并通过PCR梯度实验进一步优化模板DNA质量浓度、退火温度及退火时间，获得最佳反应条件：模板DNA质量浓度为30 ng/μL，退火温度为48℃，退火时间为20～25 s.对最佳反应体系和反应条件进行了检验，结果显示该反应体系稳定性高、重复性好
Abstract:
SSR-PCR reaction system in Carcinoscorpius rotundicauda using microsatellite primers from Tachypleus tridentatus was constructed and optimized by employing L9(34)orthogonal design with three levels and four factors,which were the concentrations of dNTPs,primers,Mg2+ and TaqDNA polymerase.A pair of microsatellite primers from Tachypleus tridentatus was used to amplify microsatellites in Carcinoscorpius rotundicauda.Finally,the optimal SSR-PCR reaction system in Carcinoscorpius rotundicauda was established in 20μL reaction volume,containing 1.5 U TaqDNA polymerases,0.16 mmol/L dNTPs,0.2 μmol/L primers and 2.0 mmol/L Mg2+.Furthermore,the optimal reaction condition with the optimal concentration(30 ng/μL) of template DNA,optimal annealing temperature(48 ℃)and annealing time(20～25 s)was obtained by gradient PCR reaction.Verification test of the optimized SSR-PCR reaction system and amplificantion procedures showed that this amplification system was stable and practicable

参考文献/References:

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更新日期/Last Update: 2014-06-28