GSM1163977: mRNA-seq Puro; Homo sapiens; RNA-Seq
Cells were subjected to 150 mJ/cm2 UV-C light. After irradiation cells were scrapped off the dishes in PBS. Cell pellets were shock frozen in liquid nitrogen and stored at –80 °C until use. After thawing on ice, cells were lysed in hypotonic gentle lysis buffer for 20 min. The cell lysate was cleared by centrifugation at 4 °C and 13,000 g for 15 min We then employed the steps of nuclease digestion, primer ligation and blotting of immunoprecipitated complexes to nitrocellulose from the iCLIP method, to minimize cloning of background RNA. Immunoprecipitation (IP) was performed with UPF1-specific antibodies. The cDNA library preparation was performed according to the iCLIP protocol (Konig et al., 2010)
GEO Accession
GSM1163977