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We are developing a workflow for using the Ampliseq Cancer Panel starting with DNA from single cells. We plan to use WGA but wonder if someone could recommend a kit. Genomiphi for example requires 10 ngs. A single cell will have about 6 pgs. Any advice?
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The following paper has a method to purify the Phi29 polymerase away from DNA contamination, which can be a problem when amplifying low concentrations of DNA. I am not sure if there is a commercial product yet.
Blainey, P. C. and S. R. Quake (2011). "Digital MDA for enumeration of total nucleic acid contamination." Nucleic Acids Research 39(4) e19. -
Qiagen has just introduced a product that is a single cell WGA.
Don't have any experience with it but might be a good starting point.Comment
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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...
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