which qiagen kit could be used in DNA recovery in ChIP assay? - (Jun/27/2008 )

Hi, everyone, I've been doing ChIP assay recently. I am wondering which Qiagen kit could be used in DNA recovery, many protocols mentioned the Qiagen PCR purification kit. It works well. How about the Qiagen gel extraction kit, can it work well too? What's the difference between PG buffer used in gel extraction and PBI buffer used in PCR purification? Or could anyone share the formula of PBI buffer or similar buffers since I may need to use a lot, and the volume provided by the kit is not enough. But I have a lot of PG buffer in the lab.

Thanks very much.

-Jason

I have been using the Qiagen PCR clean up kit which uses the spin columns. Unfortunately I have no knowledge of whats in the buffers, but I too have experienced running out of PBI buffer! Perhaps you could contact a Qiagen sales rep who may be able to pull some strings and get you a spare bottle or two of the PBI buffer.

Good luck!

Yes we also use the PCR clean up kit. We order extra PB buffer because you're gonna need a lot.

yes. i check online and qiagen does sell PB buffer separately. but can i use PB instead of PBI for ChIP? is there any difference between these two? thanks.

PBI is the same as PB, but has an indicator to check pH during the extraction. Convenient, but not necessary.

If you aren't pulling down very much DNA you might want to try the mini-elute version of the cleanup kit. It has smaller capacity but that also means less is retained on the membrane after elution which is great if you don't have much IP'd DNA. The only problem is that they expect you'll be using small volumes with the kit so they won't give you enough buffer for more than a few cleanups using standard elution volumes for ChIP. We just buy the buffers separately when we run out.

I do not think you need to use a DNA purification kit to do CHIP. With new advance in fast Chrompatin IP kit, you can do it in microplate well and finish it in 4 hours with no need to purify your DNA. There is a simple, fast chromatin IP kit available ! It is easy, reproducible doing reaction in microplate well. I tried it from {edit} and it worked very well.