Northern Blotting of siRNA Targets
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 : Shenzhen University
Other organizations : Shandong Agricultural University, University of California, Riverside
Variable analysis
- Selected siRNAs
- SiRNA expression levels
- Total RNA amount (20 μg) loaded into the gel
- Gel conditions (15% (w/v) TRIS BORATE-EDTA (TBE)–urea polyacrylamide gel)
- Electrophoresis conditions (150 V for ∼1 h)
- Transfer conditions (semi-dry method)
- Crosslinking conditions (90-min EDC-mediated chemical crosslinking at 65°C)
- Hybridization conditions (with biotin-labeled probes complementary to the siRNAs or U6 at 55°C overnight)
- Washing conditions
- Detection method (Chemiluminescent Nucleic Acid Detection Module)
- Biotin-labeled probes complementary to U6 (for normalization)
Annotations
Based on most similar protocols
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!