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My question pertains to the generation of polyclonal antibodies from synthetic peptides, such as those provided by GenScript, for use in ChIP-Seq. I recognize a potential problem in submitting a single peptide for antibody generation: that the specific protein epitope recognized by the antibody and represented by the synthetic peptide will not be exposed after formaldehyde cross-linking of the protein-DNA complex. Would it therefore be possible to obtain a polyclonal antibody by inoculating rabbits with two synthetic peptides from the same protein, such that more than one epitope may be recognized by the antibody?
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I don't think the 2-peptide strategy will work because each of the peptide will be independently displayed by the immune system. I know of a few companies that are trying this approach (with a single peptide) and are having marginal success.
If you are married to using a peptide antigen, you may want to try raising a monoclonal so you can screen for the desired properties (recognizing DNA-bound protein). Here though, the major challenge is having target to use in your screening.
Another thought would be to try a genetic immunization approach (like SDI). I've never tried them, but it looks look on paper...
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I’m not a sequencing expert. I’m a purification scientist who uses NGS to evaluate workflows my group develops. With this perspective, we think about the sample first and the NGS workflow second. The sequencer is an exceptionally honest reporter, but it can only report on what you give it, so whether you get clean, interpretable data from an NGS workflow is largely determined before you begin.
Here are nine questions we think about, in roughly the order they matter, before...06-18-2026, 07:11 AM -
Data variability is still an issue in sequencing technologies despite the advances in reproducibility and accuracy of these platforms. But the problem does not originate in the sequencing itself, but in the previous steps, before the sample reaches the sequencer.
The first step is collection, followed by preservation and sample preparation for analysis. Most scientists overlook those steps, but not being careful might just be skewing the experiment’s results.
...06-02-2026, 10:05 AM
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