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Two recent articles suggest that CRISPR/Cas9 can cause mutations in the p53 pathway, irrespective of the target sequence of the guide RNA. The data are extremely interesting since they raise the issue of unpredictable off-target effects that are not directly introduced by the CRISPR/Cas9 complex but rather selected during the genome manipulation process. A thorough analysis of the genomic integrity of cells genetically manipulated with CRISPR is, therefore, the key to successful use of the technology, as described in https://perkinelmer-appliedgenomics....w-crispr-cas9/.
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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.07-01-2026, 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...