GSM7082525: Cal51_Ran-D; Homo sapiens; RIP-Seq
Cells grown to 80% confluency in 150mm dishes were rinsed with ice-cold PBS and UV irradiated with 250 mJ/cm2, 254 nm, in ice-cold PBS using a Spectrolinker XL1500 (Spectro-UV). Cells were lysed in the dish with 1ml 1 X PXL [1 X PBS, 0.1% SDS, 0.5% deoxycholate, 0.5% Igepal] + EDTA-free Complete protease inhibitor cocktail (PIC; Roche), & lysate collected by scraping. 3' adapter NEBNext 3'SR adaptor for Illumina; /5rApp/AGA TCG GAA GAG CAC ACG TCT /3AmMO/. 5' adapter /5AmMC6/GUUCAGAGUUCUACAGUCCGACGAUCNNNNNNNNNN. RT primer 5'-AGACGTGTGCTCTTCCGA-3'. PCR amplification with a common forward primer (NEBNext SR primer for Illumina) and barcoded reverse primers for each sample (NEBNext Index primers for Illumina).