Mus musculus
source_name
cell line
extraction protocol
cells were lysed with NP-40 Lysis Buffer supplemented with cOmplete EDTA-free protease inhibitor, passed through a G26 needle before centrifugation at 20,000 g at 4 C for 10 min. The rest of supernatant was diluted 5-fold using NET-2 buffer supplemented with 10 u of Turbo DNase (Ambion) and 10 mM of MgCl2. Protein A Dynabeads that were coupled with J2 or mouse IgG2a isotype-specific control antibody were added to the supernatant and rotated at 4°C for 2 hours. Beads were washed twice with NET-2 buffer, twice with HSWB and another two times with NET-2 buffer. After all liquids were drained from the beads, RNAs were isolated from the beads using TRIzol and RNA Clean & Concentrator-5 (Zymo Research)
sample type
pulled-down RNA
molecule
immunoprecipitated RNA