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Chapters
- 00:04Introduction
- 00:42Short Tandem Target Mimic (STTM) Fragment Amplification
- 01:40STTM Purification
- 02:22T4 DNA Polymerase Reaction Mix Preparation
- 03:13Potato Virus X (PVX)-Based microRNA (miRNA) Silencing (VbMS) Vector Construction
- 04:55T4 DNA Polymerase Reaction Setup and STTM Cloning
- 07:21Potato Virus-X-Based VbMS Assay
- 10:07Results: Representative VbMS of miR165/166 in Tetraploid Potato Species
- 11:16Conclusion
We present a detailed protocol for potato virus X (PVX)-based microRNA silencing (VbMS) system to functionally characterize endogenous microRNAs (miRNAs) in potato. Target mimic (TM) molecules of miRNA of interest are integrated into the PVX vector and transiently expressed in potato to silence the target miRNA or miRNA family.
