Tweet
I'm designing some qPCR reactions to test some candidate genes before we get our RNA-seq results back. Does anyone have a good suggestion for a control gene to use for bacterial samples? I've heard that just using 16S rRNA is problematic because it is so abundant. Also, since rRNA shouldn't be in my RNA-seq reactions, I'm hesitant to use that as a control. I've seen a lot of people use recA. Is this common, or are there other suggestions?
-
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