1 reply to this topic

[MunkySpunk]

Hi,

I'm about to embark on a massive number of ChIP-chip's.

I like the chelex method, bu I've only used it for diagnostic ChIP's where the PCR product is the end result with no further processing of the IP'd DNA.

Has anyone used the chelex method for chip-chip successfully? I'm planning to use the random-priming method for total DNA amplification which requires most of the IP'd DNA to be used in a single klenow reaction followed by standard PCR.

Any thoughts? Experiences? Suggestions? Caveats?

Thanks!!

[KPDE]

MunkySpunk, on Apr 1 2010, 10:32 AM, said:

Hi,

I'm about to embark on a massive number of ChIP-chip's.

I like the chelex method, bu I've only used it for diagnostic ChIP's where the PCR product is the end result with no further processing of the IP'd DNA.

Has anyone used the chelex method for chip-chip successfully? I'm planning to use the random-priming method for total DNA amplification which requires most of the IP'd DNA to be used in a single klenow reaction followed by standard PCR.

Any thoughts? Experiences? Suggestions? Caveats?

Thanks!!

The considerations to worry about with the chelex method are that the DNA will mostly be single stranded (due to the 95C heating step) and there is no actual cleanup step in the original method (if your amplification requires purified DNA then you'll need to use mini-elute cleanup or something similar).