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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 establish a suitable feeding method for small population of Anopheles anthropophagus, to explore the influence of different preservation methods of Anopheles sinensis on total RNA extraction, and to provide the basis for providing high quality total RNA samples for correlation study. Methods The growth habit of Anopheles anthropophagus was fitted. Various measures were taken in the laboratory to improve the hatchability, larval survival rate, pupalization rate and emergence rate of Anopheles mosquitoes. Effect of Low Temperature Freezing on Total RNA Degradation. Results In the non-professional breeding room, the adoption of a simple and inexpensive method to improve the hatching rate, larval survival rate, pupalization rate and emergence rate of Anopheles anthropophagus has been greatly improved. Cryo-freezing method is better than traditional ether anesthesia method to avoid excessive degradation of specimen RNA. Conclusion A low-cost and high-success rate Anopheles anthropophagus rearing method and a low-degradation, safe and easy-to-use method for the conservation of Anopheles anthropophagus were established.