家禽血液基因组DNA的快速提取

Rapid Extraction of Genome DNA from Avian Blood

[目的]建立一种应用硅胶膜提取禽血DNA的新方法。[方法]应用硅胶膜法和传统的酚/氯仿法提取抗凝禽血基因组DNA,设计合成了一对2.7 kb的鸡卵清蛋白基因的引物,检测这2种方法提取的基因组DNA模板对聚合酶链式反应(PCR)扩增效率的影响。[结果]用硅胶膜法提取的基因组DNA产量和质量均比传统的酚/氯仿法高,将该法提取的基因组DNA作为模板能够消除抗凝剂对后续PCR反应的影响。[结论]硅胶膜法为快速提取高质量的禽血基因组DNA奠定了基础。

[Objective] To establish a novel method for extracting genome DNA from avian blood sample by silica membrane.[Method] The extraction of genome DNA from EDTA and Heparin anticoagulated chicken blood were performed by the silica membrane adsorption method and phenol-chloroform extraction method.A pair of 2.7 kb primer was designed,synthesized and used to check the genomic templates for influencing on PCR amplification efficiency by different extraction methods.[Result] The yield and quality of genomic DNA extracted by the silica membrane adsorption method was better than that extracted by the traditional phenol/chloroform extraction method.Moreover,through using the genomic DNA of silica membrane adsorption method as template of Polymerase Chain Reaction（PCR）,the influences of anticoagulants on the amplification efficiency were eliminated.[Conclusion] Silica membrane adsorption method lays the foundation for simple and rapid extraction of high quality genome DNA from avian blood.