Magna chip seq chromatin immunoprecipitation kit

To validate the regulated relationship of promoter, we conducted Chromatin immunoprecipitation experiment using Magna ChIP-Seq™ Chromatin Immunoprecipitation Kit (Millipore, Billerica, USA) following the corresponding instructions. Firstly, we conducted formaldehyde crosslinking and ultrasonic crushing of cells and impurity removal and antibody feeding. Next, we conducted precipitation and cleaning of immune complexes. All DNA samples were recycled and started PCR operation.

Magna ChIP-seq™ Chromatin Immunoprecipitation Kit (Millipore, Billerica, USA) was used for detecting protein-chromatin interactions. Briefly, the cells were fixed with formaldehyde and sonicated, and incubated with the target protein. The cross-linked DNA fragments were then released from the co-precipitated complexes, purified, and amplified by PCR. Sequences of primers amplifying the TRIM11 promoter regions were as follows: Region 1 (R1), sense: 5'- CCCAGGCTGCCCATAGAAAT-3', antisense: 5'- TTTGGGGATCGAACGGGATG-3'; Region 2 (R2), sense: 5'-GTGGGAGGTTTGGGGGACTG'-3'; antisense: 5'-CCTCAAGGGGCCTGGTTGAAA-3'.

To detect the interaction between the DNA and proteins, Chromatin immunoprecipitation (ChIP) experiment was performed with the Magna ChIP-seq™ Chromatin Immunoprecipitation Kit (Millipore, Billerica, USA) according to the instructions of manufacturer. Cross linked Chromatin DNA was sheared by ultrasonic, and incubated with E2F1 or IgG antibody and protein A/G beads over night at 4 °C. DNA was then extracted from the DNA-protein complex. Immunoprecipitated DNA was subjected to ChIP-PCR validation.