FAQs

Q:Do I have to purify my samples prior to downstream processes?
A:
For ChIP-qPCR kits, DNA clean up is not required before proceeding to qPCR.
For  ChIP-seq kits, DNA clean up is required for the sample before proceeding to sequencing.
 
Chromatrap ChIP-seq kits come complete with all reagents, columns or plates required to obtain high quality pure DNA for NGS library preparation’ or similar.
 
A:

Chromatrap kits do not use any beads throughout the entire IP assay. It’s that simple. Instead of beads, we use a porous solid state filter that has Pro-A or Pro-G bound to its surface. The porous filter drives maximum molecular mixing and immunocapture of antibody-chromatin complexes within a simple spin column. To find out more how our technology works, take a look at our Technology section.

A:

Choosing the right kit for your ChIP is incredibly important, as it affects both whether your tests work or not and the quality of the results produced. We created this handy Chromtrap ChIP kit guide to understand what you do so we can direct you to the right kit. Its interactive! 

A:
Different antibody sources have different affinities to protein A or G, check the affinity of the host species of your antibody. Protein G has the widest range of antibody affinities.
 
A:

Chromatrap® spin columns have been optimized to process between 50 and 50,000 ng of chromatin or between 1000-15million cells but chromatin loading is dependent on each application.

A:
Try substituting Chromatrap® buffers first as these have been designed to achieve the best quality of chromatin for use with the columns.
 
A:

The kits differ in the type of columns, slurry volumes and elution buffers. The qPCR kit should only be used when using cell lines and if you're only performing qPCR.

 

The ChIP-seq have a 1ml slurry volume vs 40µl for the ChIP-qPCR. This means that the ChIP-seq kit much more flexible, allowing you to load larger amounts of chromatin if required (1-50µg vs 1-10µg for the qPCR kit). The ChIP-seq kit is therefore suited for the use of primary cells and difficult cell types.

 

Note: Both qPCR and sequencing can be performed with the ChIP-seq kit but only ChIP-qPCR can be performed with the qPCR kit. With ChIP-Seq, the samples need to be purified prior to any downstream processing which is not required for the ChIP-qPCR kit. 

A:

Sonication is a simple and effective method of chromatin shearing which provides randomly fragmented chromatin. Provided the temperature is controlled during the sonication process and emulsification is avoided good quality chromatin can be obtained from most cell types using this method. Enzymatic shearing is useful if a sonicator is not available and is less disruptive to the epitopes of the protein of interest recognized by the specific antibody. Enzymatic shearing is essential when carrying out native ChIP (chromatin which has not been cross linked) as sonication can disrupts the protein/DNA complexes. Certain cell types may be resistant to lysis resulting in poor enzymatic shearing efficiency in this instance try sonication.