The Method of Eliminating the Wolbachia Endosymbiont Genomes from Insect Samples Prior to a Long-Read Sequencing
The Method of Eliminating the Wolbachia Endosymbiont Genomes from Insect Samples Prior to a Long-Read Sequencing
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在长读长测序之前从昆虫样本中消除沃尔巴克氏体内共生体基因组的方法
DOI：
10.1007/978-1-0716-2996-3_8
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Arakawa Kazuharu
中科院分区：
文献类型：
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作者：
Takasuka Keizo;Arakawa Kazuharu
当提取无脊椎动物的DNA用于长读序测序时，不仅需要足够数量和大小的DNA，而且根据物种，还必须消除内共生Wolbachiagenome的污染。这些要求变得很麻烦，特别是在小型物种中，可用于实验的个体数量有限。本章以寄生于蜘蛛的微小寄生蜂（Reclinervellus nielseni）为宿主，通过蜂蜜溶液对成虫进行抗生素给药，建立了消除Wolbachiagenomes的方法。自它们从茧中出现以来，利福平处理20天导致Wolbachiagenomes显着减少，同时保持纳米孔测序的良好DNA条件。然后通过汇集几个个体来获得足够数量的DNA。该方法可以应用于其他昆虫或无脊椎动物，可以通过实验室喂食液体食物来维持。
When extracting DNA of invertebrates for long-read sequencing, not only enough quantity and size of the DNA but, depending on the species, elimination of contamination of endosymbioticWolbachiagenome also has to be achieved. These requirements become troublesome, especially in small-sized species with a limited number of individuals available for the experiment. In this chapter, using tiny parasitoid wasps (Reclinervellus nielseni) parasitizing spiders as hosts, we developed a method of eliminating theWolbachiagenomes by means of an antibiotic administration to adult wasps via honey solution. Twenty days of rifampicin treatment since their emergence from cocoons resulted in a significant decrease in theWolbachiagenomes while keeping good DNA conditions for nanopore sequencing. An adequate quantity of DNA was then gained by pooling several individuals. The method could be applied to other insects or invertebrates that can be maintained by laboratory feeding with liquid food.