关键词: 蜜蜂, 干标本, 回软, DNA提取

Abstract: This work aims to explore the effects of preservation methods and temperature while softening specimens before drying the specimen on DNA from different parts of bees. The phenol - chloroform method was used to extract the total DNA from bee specimens preserved in different methods and from those dried bee specimens softened at different temperatures. The extracted DNA was analyzed and identified using agarose gel electrophoresis and the PCR amplification. The electrophoresis images showed that fine-quality DNA was extracted from all specimens,including specimens collected freshly,specimens soaked with alcohol, and specimens dried under natural conditions,and the quality of genomic DNA from the head and the leg of these specimens was better than that from other parts. The total extracted DNA results from dried bees softened under different bath temperatures and preserved half year showed that 65℃ was the optimal softening temperature,and the damages to bee’s DNA was the least,and the ideal part for extracting DNA was the bee’s leg. The results of orthogonal experiment also revealed that the leg and softened at 55℃ was the optimal combination for extracting DNA from the dried specimen. Furthermore,the results of PCR amplification confirmed that these DNA extracted in our experiment were used to successfully amplify mitochondrial 16S rRNA and CO I fragments. In summary,DNA extraction was explored from the three aspects：preservation methods,extracted parts and softening operations,which could provide better alternative proposals for collecting DNA from bees.

Key words: bee, dried specimens, softening, DNA extraction