title: Illumina HiSeq 2500 sequencing; RNA-Seq of A375 cells treated with tetracycline-based crosslinking probes to identify binding sites for the tetracycline analogs Col-3 and doxycycline on the human ribosome
design: RNA-Seq of A375 cells treated with tetracycline-based crosslinking probes to identify binding sites for the tetracycline analogs Col-3 and doxycycline on the human ribosome
library_construction_protocol: Following compound treatment and UV irradiation, cells were directly lysed in dishes with Ribozol reagent. All samples were subject to RNA capture/pulldown using Desthiobiotin-Streptavidin. Direct-zol RNA purification kit (Zymo Research) Total RNA was subjected to fragmentation, conjugation of desthiobiotin using \"click\" chemistry, ligation of a 3'-adapter, reverse transcription to cDNA, capture and enrichment with streptavidin beads, and PCR amplification.
attributes.tag: Experimental Factor: compound
attributes.value: doxycycline
attributes.tag: Experimental Factor: dose
attributes.value: 20
attributes.tag: Experimental Factor: irradiate
attributes.value: UV radiation 365 nm for 20 min