Protocol tips
Upstream tips |
Protocol tips |
Downstream tips |
- Spin the solution down to the bottom prior to use.
- Check if any buffers contain salt precipitates before use. If so, shake the buffer until the salts are re-dissolved. |
-For optimal preparation, the input amount should be 1 to 5 x 106 cells or 50 to 200 mg tissues.
-Avoid multiple freeze/thaw cycles. |
|
Upstream tips |
- Spin the solution down to the bottom prior to use.
- Check if any buffers contain salt precipitates before use. If so, shake the buffer until the salts are re-dissolved. |
Protocol tips |
-For optimal preparation, the input amount should be 1 to 5 x 106 cells or 50 to 200 mg tissues.
-Avoid multiple freeze/thaw cycles. |
Publication protocol
Chromatin preparation in whole bone homogenate was carried out using ChromaFlash Chromatin Extraction Kit (Base Catalog # P-2001 Epigentek, Farmingdale, NY, USA) as described by the manufacturer’s instructions. Chromatin immunoprecipitation assay was performed as per previously described41. Briefly, the immunoprecipitated DNA (through anti-H3K27ac antibody) was amplified by PCR reaction using GoTaq® Hot Start Green Master Mix (Promega, USA) and PCR was performed by using S1000™ Thermal Cycler-BIORAD. Primers were designed for the targeted gene promoter.
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