水溞RAPD反应体系的优化和建立

Establishment and Optimization of RAPD Reaction System in Daphnia

以大型溞、发头裸腹溞、蚤状溞和老年低额溞为研究材料,采用改变退火温度和循环次数等条件对RAPD扩增技术进行了优化研究.结果表明:在条件优化后(退火温度为42℃,循环次数为46次)有5条RAPD引物(A-04,A-07,B-12,C-11和C-20)在4个群体中均有扩增条带.由此表明,退火温度和循环次数的优化有利于水溞RAPD鉴定技术的建立.该研究将为水溞的遗传多样性分析奠定基础.

Four specific clones including Daphnia magna , Moina irrasa , Daphnia pulex, Simocephalus vetulus were used to optimize PCR amplification conditions to develop DNA fingerprinting technology by RAPD primers. The results demonstrated that 5 RAPD primers including A- 04, A- 07, B - 12, C - 11 and C -20 were amplify PCR products in four different populations under the optimization conditions that annealing temperature is at 42 ℃ and reaction cycle is 46. Therefore, the optimization of annealing temperature and reaction cycle numbers is beneficial to the development of RAPD technique of Daphnia. Meanwhile, this study can provide the interest intbrmation for genetic diversity of Daphnia.