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I'm having a lot of issues with NuGEN's Ovation 3'DGE and RNA-Seq library preparation kits. No matter what I do, I can't seem to generate any libraries for sequencing. Has anyone had a similar problem? When I use the mRNA Seq prep kit from Illumina and try it on the same samples I'm getting great libraries....hmmm.....frustrated....
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are you using Illumina kit to generate library itself, or your own reagents? Anyway, there is a report in Biotechniques that shows that pretreatment of ovation-generated ds cDNA with S1 nuclease increases efficiency of library generation. Their claim is that it's due to hairpins at the ends of DNA fragments that obviously would affect any end repair /adaptor ligation. S1 ligase will get rid of the ssDNA, so should solve the problem.
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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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