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  • Improved library preparatio...
    Buchbender, Andreas; Mutter, Holger; Sutandy, F.X. Reymond; Körtel, Nadine; Hänel, Heike; Busch, Anke; Ebersberger, Stefanie; König, Julian

    Methods (San Diego, Calif.), 06/2020, Letnik: 178
    Journal Article

    Individual-nucleotide resolution UV crosslinking and immunoprecipitation (iCLIP) is a state-of-the-art technology to map the RNA interaction sites of an RNA-binding protein (RBP) across the transcriptome. Here, we present the new iCLIP2 protocol that allows to obtain high-quality iCLIP libraries in a fast and efficient manner. The new protocol comprises separate adapter ligations, two cDNA amplification steps and bead-based size selection. The full procedure can be completed within four days. Our advances significantly increase the complexity of the iCLIP2 libraries, resulting in a more comprehensive representation of RBP binding sites. Overall, the methodological advances in iCLIP2 allow efficient library generation and thereby promote the versatile and flexible application of this important technology.