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This is a kind of silly question but hope somebody can help me out here. When people do NGS, do they need to spike in controls? Can someone maybe direct me to a few papers on this?
Thanks a lot!
Thanks a lot!
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I haven't heard of anyone spiking in controls, though perhaps in a few less common pipelines it is used. As far as I know it's not used for RNA-Seq, SNP calling, genome assembly or ChIP-Seq. The controls most people use are simply a control condition, e.g. untreated cells for RNA-Seq, wild type for SNP calling, different IgG pull-downs for ChIP-Seq (there was some discussion about these earlier). I'm not sure what you could use as a control for genome assembly, but it may or may not require one.
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Love this - good data definitely starts from good input, and poor input can only give relatively poor data. I particularly like the mention of Nanodrop/absorbance based methods for quantification. It's such a toss up if you'll get an accurate reading or what amounts to a randomly generated number, and a lot of library/sequencing related issues can be traced back to poor quant.Yesterday, 11:43 AM
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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... -
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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