iCLIP and iCLAP for TIA1 and TIAL1 proteins
iCLIP and iCLAP for TIA1 and TIAL1 proteins
HeLa cells grown in a 10 cm plate. Medium was replaced with ice-cold PBS buffer and cells were subjected to UV-C irradiation (100 mJ/cm2, Stratalinker 2400). Cells were then precipitated by centrifugation for 1 min at 14000 rpm and snap-frozen on dry ice. iCLIP or iCLAP protocol was followed to isolate RNA bound to TIA1 and TIAL1 proteins and amplify cDNA library as described in the manuscript. Specific barcodes were used for each RT reaction, and are specified below for each sequencing run. Random barcodes were also incoperated into cDNAs to distinguish between PCR duplication of cDNA products. The final products were obtained by PCR with Illumina pair-end sequencing primers: 5-CAAGCAGAAGACGGCATACGAGATCGGTCTCGGCATTCCTGCTGAACCGCTCTTCCGATCT-3; 5-AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCT-3
Experimental Factor: TEST
iCLAP