Department of Agronomy, Purdue University, Indiana, United States of America
Research Article
An Optimized dsRNA Production Protocol from Bacterial Cultures: A Case for Fusarium graminearum Genes
Author(s): Binod Gyawali, Houshang Alizadeh and Mohsen Mohammadi*
Double-stranded RNA (dsRNA) triggers RNA interference (RNAi), leading to the directed silencing of specific genes. This mechanism has become significant and recognized as a sustainable tool in crop disease and pest- management applications. In vivo transcription of dsRNA in bacterial cells and in vitro transcription using kits are two methods of dsRNA production at laboratory scale. Design and testing of dsRNA at laboratory-scale requires efficient dsRNA production that yields large and high-quality dsRNA in a relatively lower price. In this report, we present an optimized protocol for production of double-stranded RNA (dsRNA) from bacterial cultures, achieving significantly higher yields compared to existing laboratory methods by five-fold (on average). Post extraction enzymatic digestion experiment revealed that the extracted dsRNA is pure and contain.. View more»