Reverse crosslinking in ChIP-exo
Reverse crosslinking is a necessary step in many molecular biology techniques, such as ChIP-exo. It is necessary to revert the bonds between biomolecules in order to facilitate the purification of one of them.
In ChiP-exo, this step will separate the target DNA from the transcription factors. In order to do so, an elution buffer containing sodium dodecyl sulfate (SDS) should be added to the sample. SDS is a detergent that will denature the proteins, breaking any links between them and other biomolecules. Furthermore, the mix is incubated at 65 °C to also reverse the cross-links formed by formaldehyde.