|Register||Search||Today's Posts||Mark Forums Read|
|ChIP Chromatin Immunoprecipitation Forum Chromatin Immunoprecipitation Forum. Discuss, chat about, and post questions about the ChIP assay, in the ChIP forum. Includes troubleshooting, protocols, buffer preparation, and more.|
| ||LinkBack||Thread Tools||Display Modes|
I'm not sure what is happening with my procedure so I need your ideas if you please.
I'm working with 25x10^6 293T cells concentratred in 1mL. I need this amount to the ChIP assay.
When I go to sonicate, and I'm testing different conditions, I only get smears higher than 1000bp.
I tested different cells concentration: 12x10^6 and 5x10^6
I'm working in the maximum power p/10 seconds avoiding overheating the samples.
I didnīt test the % duty cicle, but I will...
But anyone have another idea? I need 200bp fragments.
Re: Sonication innefective?
For the 61 views until today and no answers, here is the answer:
Is the formaldehyde %.
I'm using 1%.
I tested 0,2%; 0,4%; 0,6%; 0,8%. With 0,8% i get my 200 bp bands and the CHIP worked.
Hope that this could help anyone.
|innefective , sonication|
|Thread||Thread Starter||Forum||Replies||Last Post|
|Centrifuge after sonication||halfro22||Western Blot Forum||1||10-09-2012 01:39 PM|
|Protein disruption by sonication?||naddel||ChIP Chromatin Immunoprecipitation Forum||0||07-23-2009 01:33 PM|
|Sonication||naddel||ChIP Chromatin Immunoprecipitation Forum||1||04-29-2009 05:46 PM|
|ChIP Assay Sonication Problems||IoDiNE||ChIP Chromatin Immunoprecipitation Forum||3||02-17-2009 01:12 PM|
|Sonication of tributyrin with media chang color to milky||muhammad yasir||Protocols and Methods Forum||1||04-07-2007 02:08 PM|