Tweet
Does anyone use Fluidigm C1 instrument on cells using DNASeq script and final library prep for NGS? Our lab has followed their published protocol using a control cancer cell line & we haven't got good alignment of reads across all chromosomes and have run into some other technical issues that makes us think the WGA used on the C1 isn't that good. Part of the problem is that nearly all the published papers use C1 for RNA-Seq while there are very few DNA-Seq papers out there.
-
-
Every WGA method isn't that good. -
Do you use the C1 NextGenSeq? Fluidigm gives the illustra WGA kit as their choice. When using that, it doesn't work to evenly amplify the template. Does anyone use another kit than the one Fluidigm recommends in their protocol or another process that would be better to perform DNA Seq on single cells?Comment
-
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
-
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...
Comment