Lucchetti Chiara, Genchi Marco, Venco Luigi, Bazzocchi Chiara, Kramer Laura H, Vismarra Alice
Department of Veterinary Science, University of Parma, Parasitology Unit, Parma, 43126, PR, Italy.
Clinica Veterinaria Lago Maggiore, Arona, 28041, NO, Italy.
MethodsX. 2019 Oct 31;6:2601-2605. doi: 10.1016/j.mex.2019.10.023. eCollection 2019.
, the etiologic agent of canine heartworm disease, like several other filarial nematodes, harbors the bacterial endosymbiont . To investigate metabolic and functional pathways of and individually, 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 adult , where female worms generally reach ∼25 cm in length. More importantly, adult worms of can 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 and population assessment following different treatments, an optimized protocol for co-extraction of DNA and RNA from a single sample of adult has 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 of .
犬心丝虫病的病原体，与其他几种丝虫线虫一样，携带着细菌内共生体。为了分别研究和的代谢及功能途径，以及它们之间的相互作用，转录组学和基因组分析的应用越来越普遍。虽然有几种商业试剂盒可用于单独提取DNA或RNA，但尚未描述从像成年这样的大型生物体中同时提取DNA和RNA的具体方案，成年雌虫一般长约25厘米。更重要的是，只能通过对实验感染犬进行尸检或通过对自然感染犬进行微创心脏丝虫摘除术来获得成年虫体。这使得每个单独的虫体样本都极其重要。因此，在一个旨在评估不同处理后的基因表达分析和种群评估的项目背景下，已开发出一种从成年单一样本中共提取DNA和RNA的优化方案。•使用TRIzol®试剂从大型线虫中共提取DNA/RNA的优化方法。•能够最大程度地利用作为独特样本的成年虫体。
