iCLIP-seq
iCLIP-seq (individual-nucleotide resolution UV crosslinking and immunoprecipitation) is a method used to identify protein-RNA interactions. This method uses ultraviolet light to covalently bind proteins and RNA molecules, with less background than RNA immunoprecipitation. As with all other methods of CLIP, iCLIP to connect the purification of proteins-RNA compounds is very strict to use immune precipitation, SDS-page, and then transferred to nitrocellulose membrane, finally remove radioactive marker protein-RNA complexes, RNA is released by using protease treatment in RNA crosslinking sites leave one or two amino acids.
Fig 1 Overview of the iCLIP experiment[1].
Application of iCLIP-Seq:
- Analyze the mechanism of RNA-protein interaction
- Interactions between incRNAs or miRNAs and RNA were analyzed to find potential RNA targets
- Genome-wide RNA screening
- Find RNA screening targets
The basic process of iCLIP-Seq:
Advantages of our iCLIP-Seq service:
- Nucleotide resolution of protein binding sites
- Avoid using nuclease
- High throughput amplification has high detection accuracy
- Low background, clear results, reduce PCR experiment time
- The improved ICLIP-Seq can reduce the reproducibility bias in non-linear PCR amplification
Workflow of Lyophilization service in Profacgen:
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Reference:
[1] Busch Anke, Brüggemann Mirko, Ebersberger Stefanie, et al. iCLIP data analysis: A complete pipeline from sequencing reads to RBP binding sites [J]. Methods, 2020, 178:49-62.