Short Tandem Target Mimic (STTM) Fragment Amplification
1:40
STTM Purification
2:22
T4 DNA Polymerase Reaction Mix Preparation
3:13
Potato Virus X (PVX)-Based microRNA (miRNA) Silencing (VbMS) Vector Construction
4:55
T4 DNA Polymerase Reaction Setup and STTM Cloning
7:21
Potato Virus-X-Based VbMS Assay
10:07
Results: Representative VbMS of miR165/166 in Tetraploid Potato Species
11:16
Conclusion
Transkript
The traditional approach to analyzing the function of microRNAs in plants primarily relies on the generation of stable knockdown plants. This virus-based microRNA silencing system does not require tedious and time-consuming transformation. Virus-b
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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.