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ChIPs cloning


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#1 Cati

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Posted 26 January 2005 - 06:01 AM

Hi, everybody!
I did ChIP on Tera2 cell line with all possible controls. :angry:
Now, I want to ligate adapters to ChIP DNA.
Could you help me, and tell smth. :)  about it, and about some problems in this work?

#2 chromatin

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Posted 09 April 2009 - 05:03 AM

View PostCati, on Jan 26 2005, 09:01 AM, said:

Hi, everybody!
I did ChIP on Tera2 cell line with all possible controls. :P
Now, I want to ligate adapters to ChIP DNA.
Could you help me, and tell smth. :rolleyes:  about it, and about some problems in this work?

How did you fragment the DNA? Sonication, Micrococcal nuclease? You need an end repair enzyme kit or individual enzymes which makes ends ligatable. So you need to fill the overhangs or get rid of them and add phosphates so that you can ligate them to the linkers that has no phosphates.
If you want commercial kit use the following, in our hands it was the most efficient one out of several we tested.
http://www.lucigen.c...64ab46350d095ff
It is very easy good luck.




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