Department of Nematology, University of California, Riverside, CA, USA.
Department of Developmental and Cell Biology, Center for Complex Biological Systems, University of California, Irvine, CA, USA.
Methods Mol Biol. 2021;2170:79-99. doi: 10.1007/978-1-0716-0743-5_6.
The advancement of transcriptomic studies in plant parasitic nematodes will greatly benefit from the development of single-nematode RNA-seq methods. Since many plant parasitic nematodes are obligate parasites, it is often difficult to efficiently obtain sufficient amounts of nematodes for transcriptomic studies. Here we have adapted SMART-Seq2 for single-nematode RNA-seq requiring only an individual nematode for a sample replicate. This protocol provides a detailed step-by-step procedure of the RNA-seq workflow starting from lysis of the nematode to quantification of transcripts using a user-friendly online platform.
植物寄生线虫转录组研究的进展将极大地受益于单线虫 RNA-seq 方法的发展。由于许多植物寄生线虫是专性寄生虫，因此通常很难有效地获得足够数量的线虫进行转录组研究。在这里，我们已经对 SMART-Seq2 进行了改编，使其适用于单线虫 RNA-seq，每个样本只需一个线虫即可进行重复。本方案提供了从线虫裂解到使用用户友好的在线平台定量转录物的 RNA-seq 工作流程的详细分步程序。
