双条拂粉蚧基因组DNA提取方法的比较和优化  

Comparison and optimization of methods for genomic DNA extraction from Ferrisia virgata Cockerell

作　　者：王宇航 孟秀利 黄山春[2] 林兆威 宋薇薇[2] 唐庆华[2] 覃伟权[2] WANG Yuhang;MENG Xiuli;HUANG Shanchun;LIN Zhaowei;SONG Weiwei;TANG Qinghua;QIN Weiquan(School of Plant Protection,Hainan University,Haikou,Hainan 570228,China;Coconut Research Institute,Chinese Academy of Tropical Agricultural Sciences,Wenchang,Hainan 571339,China)

机构地区：[1]海南大学植物保护学院,海口570228 [2]中国热带农业科学院椰子研究所/海南省槟榔产业工程研究中心,海南文昌571339

出　　处：《热带生物学报》2023年第6期636-641,共6页Journal of Tropical Biology

基　　金：海南省重点研发项目(ZDYF2022XDNY208);海南省自然科学基金项目(321QN346);海南省院士创新平台科研专项(YSPTZX202138)。

摘　　要：为了提高双条拂粉蚧(Ferrisia virgata Cockerell)昆虫基因组DNA的提取浓度和质量,以当天采集活体和经95%酒精-20℃冷藏浸泡7 d后的双条拂粉蚧(Ferrisia virgata Cockerell)成虫为实验材料,采用2种试剂盒、3种前处理研磨方法进行组合的方式,设置1头、3头、5头、7头和10头5个梯度,提取不同头数的粉蚧基因组DNA,并对提取的基因组DNA浓度和OD比值(A260/A280)进行了测定和评价。结果表明,以活体双条拂粉蚧为材料,使用TIANGEN试剂盒,采用研磨杵+研磨仪作为前处理,提取昆虫数为10头时,双条拂粉蚧基因组DNA浓度较高、质量较好,其OD比值(A260/A280)为1.80~1.90,电泳条带清晰;与其他组合的OD比值(1.95~2.20)相比所含杂质和污染最少。In order to improve the concentration and quality of genomic DNA extracted from the striped mealybug(Ferrisia virgata),the adults of the striped mealybug were collected on the same day and soaked in 95%alcohol for 7 days,and their genomic DNAs were extracted from a gradient of 1,3,5,7 and 10 adults by using different combinations of two kits and three pre-treatment grinding methods.The concentration and OD value of the extracted genomic DNAs(A260/A280)were estimated and evaluated.The results showed that when 10 adults were used for extraction under the combination of TIANGEN kit and grinding pestle+grinding instrument as pretreatment,the concentration and quality of the genomic DNAs of the striped mealybug were high;the OD value(A260/A280)was 1.80-1.90;the gel band was clear.Compared with other combinations(1.95-2.20),this combination extracted genomic DNA with the least impurities and contamination.

关 键 词：双条拂粉蚧 基因组DNA 试剂盒法 研磨方法 

分 类 号：Q966[生物学—昆虫学]