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Input reverse cross linking

input reverse cross link

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#1 oxygene8

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Posted 21 April 2012 - 03:25 AM

I am performing a ChIP assay using the R&D kit. According to the manufacturers protocol, the samples (antibody IP and IgG IP) are washed after the antibody incubation and then boiled with a Chelating Resin solution for 10 minutes. This basically is the final step before DNA purification using a kit. I am assuming this boiling step involves reverse cross linking for the samples. My question is, how do I reverse cross link the input DNA that I have set aside previously? The kit nowhere says anything about the processing of input DNA. Can I add Chelating Resin solution to the input and treat likewise or do I follow a traditional 65 degrees treatment (though I am reluctant to use different methods for the input and samples).

Any help or experience with a such a problem will be welcome....

#2 KPDE

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Posted 21 April 2012 - 02:18 PM

In my experience, boiling in either chelex or high pH Tris-EDTA buffers (they're essentially equivalent) is sufficient for reversal of crosslinking of input samples.

Edited by KPDE, 21 April 2012 - 02:19 PM.






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