中华鳖群体DNA指纹分析中的RAPD技术优化

Optimization of RAPD technique of fingerprinting analysis of Pelodiscus sinensis populations

以中华鳖(Pelodiscus Sinensis)4种不同区域代表性群体黄河鳖、太湖鳖、台湾鳖和日本鳖为研究材料,采用不同反应条件(如退火温度和循环次数等)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38℃,循环次数为40次)有20个RAPD引物在4个群体中均有扩增条带,其中随机引物S105能将黄河鳖群体和太湖鳖、台湾鳖、日本鳖群体区分开;随机引物S327能将日本鳖群体鉴别出来;而随机引物S474可将台湾鳖群体与其他群体分开。由此表明,退火温度和循环次数的优化有利于中华鳖RAPD鉴定技术的建立。本研究旨在为中华鳖的种质鉴定与保护提供技术支撑。

Four populations of Pelodiscus sinensis from Yellow River,Taihu Lake,Taiwan and Japan were used to optimize PCR amplification conditions to develop DNA fingerprinting technology by RAPD primers.The results showed 20 RAPD primers could amplify PCR products in four different poplations under the optimization conditions.The result also demonstrated that PCR products amplified by RAPD primer S105 in sinensis population Yellow River were different from those in other three populations,while RAPD primers S327 and S474 could differeniate Japan and Taiwan populations from other three populations,respectively.Therefore,the optimization of annealling temperature and cycles numbers is beneficial to the development of RAPD technique of P.sinensis.Meanwhile,the reasonablity and its affecting factors of RAPD technique was also briefly discussed in this paper,which can provide the interest information for similar researches.