What is the maximum length of DNA we can use in a pull-down assay?

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Prem Subramaniam

Do you mean chromatin immunoprecipitation? Can you please specify the problem?

Rishi kumar Jaiswal

Actually i want to do DNA pull down assay as well as chromatin immunoprecipitation so for DNA pull down assay what should be the proper length of the DNA.

By DNA pull down you mean like tagged-DNA that you will pull down with beads?

The length is a function of the binding site - for binding your protein with high enough affinity. Also, depending on whether you are looking for one protein or a complex, and how much DNA the protein occupies the size can vary between 30 bp - 300bp. I have seen studies with both. The 300-500 bP probes can pull down complexes, if one was looking for that.

Attached is a good protocol paper, that may help you.

Good luck!

Thanks alot Prem subramaniam sir.