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Hello tramelliwe I can't really speak on whether the two peaks are common, but I can say that after doing a lot of amplicon sequencing I've had a lot of off-target amplification. Depending on what you're sequencing it can be expected and it happens even more with degenerate primers. Have you looked to see if any of your primers match up with that region? I can also check in with some friends that have more experience with this type of work. -
Thank you for your answer Ben3. Maybe I should clarify that my amplicon covers both peaks. I did check the specificity, but even with multiple mismatches, my primers do not bind other places on that locus. I did not do the library prep myself, maybe at that stage the fragments covering that zone were lost somehow?
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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 -
With the launch of new single-cell sequencing platforms in 2026, the field stands at an exciting inflection point. This article surveys the most impactful advances in the field and discusses how they’re reshaping research in cancer, immunology, and beyond.
Introduction
Single-cell sequencing technologies have undergone remarkable advances over the past decade, transitioning from low-throughput experimental approaches to highly scalable platforms capable of...05-22-2026, 06:42 AM
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