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Low DNA yield from neutrophils

Low DNA yield from neutrophils - ChIP Chromatin Immunoprecipitation Forum

Low DNA yield from neutrophils - Chromatin Immunoprecipitation Forum. Discuss, chat about, and post questions about the ChIP assay, in the ChIP forum. Includes troubleshooting, protocols, buffer preparation, and more.


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Old 10-31-2011, 06:59 PM
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Default Low DNA yield from neutrophils



Hello,
I am using the Life Technologies MAGnify ChIP kit with 2 X10^6 freshly isolated human neutrophils. About 50% of the time, I get a decent smear on my agarose gel at the right size, and 50% of the time the smear is very faint. I have optimized the shearing conditions to get a population of fragments of the desired size but am struggling to understand why the yield of DNA is so variable.

I cross-link for 10 minutes with 1% formadlehyde and am using the Covaris S2 for 12 minutes shear consistently with my cells.

Any guidance about what may affect the yield of DNA would be greatly appreciated.
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