title: Illumina HiSeq 4000 sequencing; iCLIP of Dazl in mouse embryonic stem cells
library_construction_protocol: Mouse embryonic stem cells were UV cross linked and lysed in 50 mM TrisвЂ“HCl, pH 7.4, 100 mM NaCl, 1% Igepal CA-630, 0.1% SDS, 0.5% sodium deoxycholate and ROCHE c0mplete protease inhibitor cocktail (1x) RNA cross-linked to DAZL was immunoprecipitated and PAGE was performed on protein-RNA complexes. After transfer of the protein-RNA complexes to nitrocellulose (wet blotting) the complexes were digested with Proteinase K and the liberated RNA-peptide complexes purified using PCA.  Library construction was carried out according to the iCLIP protocol described elsewhere [1]. RNA was ligated with a IRL3 3'adapter, circularised and reverse transcribed using barcoded RT primers.  [1] Blazquez L et al. Mol Cell 2018. doi: 10.1016/j.molcel.2018.09.033