We obtained the sequences of the RNAi pathway core components in the genome of P. equestris through a local BLASTx search with corresponding sequences of Oryza sativa as queries (Cai et al., 2015 (link); Kapoor et al., 2008 (link); Niu et al., 2016 (link)). Furthermore, we used the sRNA target analysis server psRNATarget (Dai and Zhao, 2011 (link)) to predict putative target genes of P. equestris by CymMV- and ORSV-derived siRNAs. We then investigated the transcript levels of these core components and target genes by RT-qPCR assay on a Quant-Studio 6 Flex Real-Time PCR System (Life Technologies, Carlsbad, CA, USA) using the SYBR Green PCR Master-Mix kit (Promega, Madison, WI, USA) with primers (Table 1 ) as described previously (Lan et al., 2015 (link), 2018a (link)).
Protocol detail
Identification and Characterization of RNAi Pathway in Phalaenopsis
Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link:
Access Free Full Text.
Corresponding Organization :
Other organizations : Zhangzhou Normal University
Top 5 similar protocols
Protocol cited in 1 other protocol
Variable analysis
independent variables
- RNAi pathway core components in the genome of P. equestris obtained through local BLASTx search with corresponding sequences of Oryza sativa as queries
- Transcript levels of RNAi pathway core components and target genes of P. equestris measured by RT-qPCR
- Primers used for RT-qPCR as described previously (Lan et al., 2015, 2018a)
- SYBR Green PCR Master-Mix kit (Promega) used for RT-qPCR
- Quant-Studio 6 Flex Real-Time PCR System (Life Technologies) used for RT-qPCR
- No positive or negative controls were explicitly mentioned in the input protocol.
Annotations
Based on most similar protocols
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
Sign up for free or login to display all annotations
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!