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Our lab is making a library from yeast total RNA. We have tried NuGen kit, and got huge rRNA peaks in the samples. We then tried Illumina's RNA-Seq and got no cDNA at all. Then we tried Ambion's mRNA purist to pull out just the Poly A, and are still getting significant rRNA carryover. Has anyone else had such problems with yeast RNA samples?
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Hi, I have been having this kind of problem as well isolating human polyA mRNA using Qiagen oligotex. Have you tried doing two rounds of purification? One of the Qiagen boffins has suggested that the rRNA contamination after purification might have something to with rRNA associated with translation of your mRNA... Do you have some pics of your bioanalyzer traces that show you your contam?
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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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