GSM3034500
GSM3034500: ChIP-Input-rep1; Mus musculus; ChIP-Seq
Hind limbs muscles were collected and placed in tube with 4 ml of cold buffer A (300 mM sucrose, 10 mM NaCl, 1.5 mM MgCl2, 15 mM Tris HCl pH 8.0 and protease inhibitor cocktail (Sigma)). The tissue was first mince with scissor and then transfer in a dounce homogenizer where 50 strokes of the loose pestle was slowly applied.The extract was then passed through a 100 µm cell strainer to remove myofibrils debris and placed back into a clean cold dounce homogenizer. To release nuclei, NP-40 was added to a final concentration of 0.5% and incubated for 15 min at 4°C. An additional 50 strokes with the tight pestle is then applied. Lastly extract was further clarified by passing through a 40 µm cell strainer, centrifuged at 2000 g for 10 min at 4°C and was washed in cold buffer A. Nuclei were resuspended in a small volume of buffer A before ChIP or immunoblot analysis. Libraries were prepared using the Diagenode MicroPlex Library Preparation kit v2.
GEO
GSM3034500
GEO Accession
GSM3034500