嗜人按蚊种群饲养及标本保存方法对总RNA提取影响的研究

Investigation into the impact of methods in raising and keeping of Anopheles anthropophagus on the extraction of total RNA

目的建立适合嗜人按蚊小型种群饲养方法,探讨按蚊标本不同保存方法对总RNA提取的影响,为进行相关性研究提供高质量总RNA标本提供依据。方法拟合嗜人按蚊的生长习性,在实验室采用多种针对措施以提高按蚊的孵化率、幼虫生存率、蛹化率和羽化率;采用总RNA变性胶凝胶电泳分析乙醚麻醉和低温速冻法对总RNA降解的影响。结果在非专业饲养室中采用简便低廉的改良方法大大提高了嗜人按蚊的孵化率、幼虫生存率、蛹化率和羽化率。低温速冻法较传统的乙醚麻醉法更好得避免了标本RNA的过多降解。结论建立了一种低成本、高成功率的嗜人按蚊饲养方法和一种低降解、安全简便的差异分析用嗜人按蚊标本的保存方法。

Objective To explore the laboratory breeding of An. anthropophagus and extraction of total RNA for different gene expression analysis. Methods Several methods were used for enhancement of the rates of hatching, survival, pupation and eclosion of An. anthropophagus. The impact of aether anaesthesia and low temperature freezing methods on the degradation of An. anthropophagus RNA was also analyzed, Results The rates of hatching, survival, pupation and eelosion of An. anthropophagus were greatly enhanced after use of the modified methods. The low temperature freezing technique was superior to that of the aether anaesthesia methods to have prevented the excessive degradation of RNA, Conclusion A technique for breeding An. anthropophagus with low cost, high success and a differentiating method for preserving An. anthropophagus samples with low degreadation, safety and simpliety have been established.