GSM8001566: RIP-Ctrl-Input; Mus musculus; RIP-Seq
The cultured cells were homogenized in polysome lysis buffer,and proteins from this supernatant were incubated with Magnetic Dynabeads pre-coated antibody. A portion of the supernatant was reserved as 'input'. The samples were washed and treated with DNase I and proteinase K to eliminate DNA and proteins. The immunoprecipitated RNA was harvested using Trizol (Takara). and 0.1μg of RNA was used for the construction of sequencing libraries for RIP-seq. Immunoprecipitated RNA or Input was used for library construction with NEB Next® Ultra™ RNA Library Prep Kit (New England Biolabs) following manufacturer's protocols.