斜带髭鲷高质量DNA提取及AFLP分析体系建立

目的:提取斜带髭鲷高质量基因组DNA并建立适用于AFLP分析的体系。方法:以斜带髭鲷(Hapalogenys nitens)为材料提取高质量基因组DNA,并进行AFLP扩增。对AFLP分析体系中几个关键环节(基因组双酶切、连接、预扩增及选择性扩增、变性聚丙烯酰胺凝胶电泳和银染)做了优化。结果:提取的斜带髭鲷全基因组DNA纯度高,无拖尾。采用9组选择性引物共检测出350个不同的扩增位点,其中多态位点为175个,多态性比例为49.58%。聚丙烯酰胺凝胶电泳图像染色均匀,条带清晰且无背景干扰。结论:该分析体系的建立为斜带髭鲷的群体遗传多态性、种质资源、遗传图谱、遗传育种等研究奠定基础。

Extraction of High - quality DNA and Establishment of AFLP Analysis System for Hapalogenys nitens

Objective:To extract high-quality genomic DNA and establish a suitable for amplified fragment length polymorphim(AFLP) analysis system of Hapalogenys nitens.Method:We used AFLP technique to carry out the high-quality DNA which had extracted,and optimized several key procedures of AFLP including enzyme-digested reaction,ligation,pre-amplification selective amplification,PAGE and silver-stainin.Result:The genome DNA extracted from Hapalogenys nitens was high purity and had little trailing.9 selective primers amplified a total of 350 sites,of which polymorphic loci was 175,proportion of polymorphic loci is 49.58%,polyacrylamide gels is clear and dyeing uniformity without background interference.Conclusion:The establishment and optimize of AFLP analysis system in this paper would lay a groundwork for the research of Hapalogenys nitens genetic diversity,germplasm resource,construction of genetic linkage map,genetic breedin and so on.