Transient Assays for the Analysis of miRNA Processing and Function
Transient assays provide a convenient alternative to stable transformation. For small RNA analysis in plants, the most widely used method, commonly named agroinfiltration, makes use of Agrobacterium tumefaciens to deliver transgenes into leaf cells of Nicotiana benthamiana. Compared to the generation of stably transformed plants, agroinfiltration is more rapid, and samples can be analyzed a few days after inoculation. Agroinfiltration has been used successfully in many different applications, including the analysis of small RNAs. We describe here a protocol for analysis of miRNA processing using agroinfiltration of N. benthamiana leaves.
Key wordsAgroinfiltration Transient assay miRNA processing Small RNA blot
We would like to thank Dr. Jia-Wei Wang for discussion and Heike Wollmann for discussion and suggestions in the preparation of this chapter. FFF is supported by DAAD. Work on small RNAs in the Weigel laboratory is supported by European Community FP6 IP SIROCCO (contract LSHG-CT-2006-037900) and by the Max Planck Society.
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