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标题：Optimized protocol for DNA/RNA co-extraction from adults of Dirofilaria immitis
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作者：Chiara Lucchetti ; Marco Genchi ; Luigi Venco 等
期刊名称：MethodsX
印刷版ISSN：2215-0161
电子版ISSN：2215-0161
出版年度：2019
卷号：6
页码：2601-2605
DOI：10.1016/j.mex.2019.10.023
语种：English
出版社：Elsevier
摘要：Graphical abstractDisplay OmittedAbstractDirofilaria immitis, the etiologic agent of canine heartworm disease, like several other filarial nematodes, harbors the bacterial endosymbiontWolbachia. To investigate metabolic and functional pathways ofD. immitisandWolbachiaindividually, along with their interactions, the use of both transcriptomic and genome analysis has becoming increasingly popular. Although several commercial kits are available for the single extraction of either DNA or RNA, no specific protocol has been described for simultaneous extraction of DNA and RNA from such a large organism like an adultD. immitis, where female worms generally reach ∼25 cm in length. More importantly, adult worms ofD. immitiscan only be obtained either through necropsy of experimentally infected dogs or by minimally-invasive surgical heartworm removal of naturally infected dogs. This makes each individual worm sample extremely important. Thus, in the context of a project aimed at the evaluation of both gene expression analysis andWolbachiapopulation assessment following different treatments, an optimized protocol for co-extraction of DNA and RNA from a single sample of adultD. immitishas been developed.•An optimized method for DNA/RNA co-extraction from large size nematodes using TRIzol® reagent.•Allows maximum exploitation of unique samples as adults ofD. immitis.
关键词：TRIzol1;RNA extraction;DNA extraction;Adult worms