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I have measured the samples after the PCR and they had good measurement especially with gene B. After the beads some of them their concentration even increased. The PCR is a custom panel primers that are ready designed and have been tested before and worked very well.
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If you can post the concentrations before and after clean up.Comment
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Also you said your pre/post clean up volumes were the same you're eluting into 25ul. 5ul more than the PCR reaction. This would account for a ng/ul concentration difference but not total. The minimum suggested volume for SPRI elution is also 30ul.Comment
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For example one of the samples was 52 ng/ul after PCR then for gene A after purification it changed to be 18,64 ng/ul. another sample was 22 ng/ul after AMPURe became 33,4 ng/ul however when I repeated the measurement now it is 8,92 after AmPure. The other problem is that the differences between the concentration in different samples is so variable between samples and with different genes. Example sample 9 is 17.38 ng/ul with gene A and 1.542 ng/ul in gene B. Could I still pool them? or run them in different chips?Comment
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Your ng/ul will be different pre and post clean up because of the volume difference. You have a 20ul PCR reaction and a 25ul clean up elution.
PCR won't amplify the same with different primers!Comment
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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.
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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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