Novus has recently released six new primer sets designed for use in real-time PCR DNA amplification of housekeeping, silent, or heterochromatin associated proteins. Specifically, the primers are for GAPDH, RPL30, MyoD1, AFM, alpha Satellite, and Satellite 2. Each primer set has been thoroughly tested and validated for chromatin immunoprecipitation using our ChIP protocol, and the results from these characterizations are available on the product datasheets.
For QC testing, chromatin immunoprecipitation was performed with one million HeLa crosslinked cells. After immunoprecipitation with either the antibody of interest or a no antibody (No Ab) control, the crosslinks were reversed and the DNA was purified. This DNA was then amplified using quantitative real-time PCR with SYBR green dye. Six different primer sets were used. GAPDH and RPL30 were chosen to represent actively transcribed genes. MyoD1 and AFM were used to represent genes that are normally silent or inactive. Finally, alpha Satellite repeats and Satellite 2 repeats were used to amplify heterochromatin associated with centromeric regions of chromosomes. All numbers represent values compared to the total, non-precipitated, input which is equal to one.
