RNA degradation of DLD-1 cells under hypoxic stress

In this work, we determined RNA degradation rate of whole transcriptome in DLD1 cells, human colorectal adenocarcinoma cells, by employing BRIC-seq (5'-bromo-uridine (BrU) immunoprecipitation chase-deep sequencing analysis) method. The cells were cultered under 1% O2 condition for overnight, followed by labeled with 150 micro molar BrU for 24h. Then medium was replaced with normal growth medium, DMEM containing 10 FBS and antibiotics, to wash out BrU. After changing medium, cells were collected and total RNAs labeled with BrU were isolated by using RNAiso Plus reagent. BrU labeled RNAs were isolated by specific mouse monoclonal antibody against BrU, clone 2B1. Finally, isolated BrU labeled RNAs were analyzed by illumina platform.