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|The ChIP assay was performed using the imprint chromatin immunoprecipitation kit (CHP1, Sigma-Aldrich) following the manufacturer's instructions. The sonicated DNA was incubated with anti-IgG (Millipore, Milford, MA, USA) as a negative control. The purified DNA was amplified using specifically designed real-time ChIP PCR primers spanning the region −300 to +1000 of LY6K.|
|-Add protease inhibitors to all lysis solutions before use.|
|-Keep cells on ice between the rounds of homogenisations.
- Increase or decrease the homogenization step to maximize the yield of nuclei depending on cell line.
-Do not place ethidium bromide in the agarose gel or the electrophoresis buffer, because of the presence of SDS.
|-It is highly critical that the chromatin is of appropriate size and concentration.|
|-It is important to keep the tissue cold to avoid protein degradation.
-Use fresh formaldehyde that is not past the manufacturer's expiration date.
|-Once in solution, store 1M DTT at -20°C.|