Illumina HiSeq 2500 sequencing; hiCLIP analysis of RNA duplexes bound by STAU1 in HEK293 cells
hiCLIP analysis of RNA duplexes bound by STAU1 in HEK293 cells
A FlpIn293 cell line expressing 3xFLAG-STAU1 was subjected to UV-C irradiation before collection. hiCLIP or iCLIP protocol was followed to isolate RNA bound to 3XFLAG-STAU1 protein and amplify cDNA library as described in the manuscript. Specific barcodes were used for each RT reaction, and are specified in the manuscript. Random barcodes were also incorporated into cDNAs to distinguish between PCR duplication of cDNA products. The final products were obtained by PCR with Illumina paired-end sequencing primers: 5-CAAGCAGAAGACGGCATACGAGATCGGTCTCGGCATTCCTGCTGAACCGCTCTTCCGATCT-3; 5-AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCT-3
Experimental Factor: immunoprecipitate
2
LIBRARY_STRAND
first strand