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Related Experiment Video

Updated: Jan 5, 2026

iCLIP - Transcriptome-wide Mapping of Protein-RNA Interactions with Individual Nucleotide Resolution
10:45

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Published on: April 30, 2011

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Improved library preparation with the new iCLIP2 protocol.

Andreas Buchbender1, Holger Mutter1, F X Reymond Sutandy2

  • 1Institute of Molecular Biology (IMB), Ackermannweg 4, 55128 Mainz, Germany.

Methods (San Diego, Calif.)
|October 15, 2019
PubMed
Summary
This summary is machine-generated.

The new iCLIP2 protocol offers a faster, more efficient method for mapping RNA-binding protein interactions. This enhanced technique improves RNA interaction site analysis across the transcriptome.

Keywords:
CLIPHigh-throughput sequencingProtein-RNA interactionRNA-binding proteinUV crosslinkingiCLIP

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Area of Science:

  • Molecular Biology
  • Genomics
  • Biochemistry

Background:

  • Individual-nucleotide resolution UV crosslinking and immunoprecipitation (iCLIP) is a key technology for mapping RNA-binding protein (RBP) interactions.
  • Existing iCLIP protocols can be time-consuming and complex.

Purpose of the Study:

  • To introduce an improved iCLIP protocol, termed iCLIP2.
  • To enhance the speed, efficiency, and quality of iCLIP library generation.

Main Methods:

  • The iCLIP2 protocol involves separate adapter ligations.
  • It includes two complementary DNA (cDNA) amplification steps.
  • Bead-based size selection is utilized for library purification.

Main Results:

  • The iCLIP2 protocol can be completed within four days.
  • It generates high-quality iCLIP libraries.
  • The protocol increases library complexity for a more comprehensive RBP binding site map.

Conclusions:

  • iCLIP2 offers methodological advances for efficient library generation.
  • The protocol promotes versatile and flexible application of iCLIP technology.
  • This facilitates a deeper understanding of RBP-transcriptome interactions.