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Has anyone run the new v3 flowcells and reagents on the HiSeq2000? We need to purchase more reagents but I am hesitant to switch over without feed back from other people.
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We haven't but will be soon. As I understand it, V3 will soon be the only option from Illumina. I actually have a question about how much library to load on the V3 kits to get in the range of 800k clusters/mm^2. On the V2 flow cells, we get about 600K with 9pm loaded. We titrated the load on the V2 lanes and the resulting cluster densities increased linearly with the load, to a point. In moving to a larger lane, I don't think the same would be true. Does anyone have experience with this? -
We'll be transitioning to v3 soon as well. Our FAS suggested starting at 12pM.Comment
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Note the optics of your HiSeq have to be recalibrated before running v3 flowcells and reagents. The Y coordinates in the config file needs updating by using a beaded flow cell by your service engineer.Comment
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Something to make a note of: The quality table for the new, v3 kits has a range from “B” to “m" per Illumina tech support.Comment
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So now the highest quality value is 45? What was it previously?
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PhillipComment
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It was 40 (http://en.wikipedia.org/wiki/FASTQ_format).Originally posted by pmiguel View PostComment
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Well the unavoidable cheap shot at Illumina is: kind of like having an amplifier that goes up to "11"?
Looks like the highest ordinal for a printable ascii char is "62" for "~". Roughly one error in 4 million base calls. For a 33 offset encoding you can reach a quality score of "93". One error in 8 trillion base calls.
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PhillipComment
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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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