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
Transcript
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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Apresentamos um protocolo detalhado para o sistema de silenciamento de microRNA baseado em vírus de batata X (PVX) para caracterizar funcionalmente microRNAs endógenas (miRNAs) na batata. As moléculas de mímica alvo (TM) de miRNA de interesse são integradas ao vetor PVX e transitavelmente expressas na batata para silenciar a família miRNA ou miRNA alvo.