Abstract
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.
Key words
- RNA-seq
- Nematode
- Smart-seq2
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Acknowledgments
This work is supported in part by NIH NIAID R21 AI142121 to A. Dillman.
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Chang, D., Serra, L., Lu, D., Mortazavi, A., Dillman, A. (2021). A Revised Adaptation of the Smart-Seq2 Protocol for Single-Nematode RNA-Seq. In: Jin, H., Kaloshian, I. (eds) RNA Abundance Analysis . Methods in Molecular Biology, vol 2170. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0743-5_6
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DOI: https://doi.org/10.1007/978-1-0716-0743-5_6
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