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I am trying to making Illumina sequencing library using transposon based method to add adaptor. However, instead of using the adaptor used in Nexera kits, I would like to use the Y-shaped adaptor. Thus, the gap between the bottom stand of adaptor and bottom strand of genomic DNA need to be repaired. I want to find a method to do gap filling, however, I do not want the DNA to the 3' of the gap (i.e., bottom strand of the adaptor) to be removed by 5'-3' exonuclease activity of DNA Pol I and then re-synthesized. Can I use Klenow fragment here? It is also strange that no one seems to use a Y-shaped adaptor in transposon based library prep. Is there any technical difficulties here?
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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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