ChIP Mouse - Gonadotrope cell lines

DNA-protein interactions are studied by using ChIP. The basic steps in this technique are crosslinking, sonication, immunoprecipitation, and analysis of the immunoprecipitated DNA. During ChIP, if chromatin is under-fragmented or fragments are too large which can lead to the increased background and lower resolution. Shorter cross-linking times (5-10 min) and/or lower formaldehyde concentrations (<1%) may improve shearing efficiency. If Chromatin is over-fragmented, then optimize shearing conditions for each cell type to improve ChIP efficiency. Over-sonication of chromatin may disrupt chromatin integrity and denature antibody epitopes. If you do not see any product or very little product in the input PCR reactions, add 5–10 μg chromatin per IP.

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Found 1 matching solution for this experiment

TruSeq ChIP Library Preparation Kit

Illumina

Upstream tips	Protocol tips	Downstream tips
-Remove End Repair Mix and Resuspension Buffer from -15°C to -25°C storage and thaw them at room temperature.	-5–10 ng ChIP-enriched, fragmented input DNA is recommended. -Carefully collect ChIP DNA samples to make sure that they are free of contaminants.

Upstream tips
-Remove End Repair Mix and Resuspension Buffer from -15°C to -25°C storage and thaw them at room temperature.
Protocol tips
-5–10 ng ChIP-enriched, fragmented input DNA is recommended. -Carefully collect ChIP DNA samples to make sure that they are free of contaminants.

Manufacturer protocol Publication protocol 1 Relevant paper

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