Ah, that’s a good point. We should probably do a full costing.
While the AffiGEN kit is more expensive per sample, I’d be tempted to use it first, as it covers everything required to check the DIYNAFLUOR and/or Open Colourimeter Plus in a manner that is less open to experimental error. Then I’d also use it to compare my results with DIYNAFLOUR low-cost protocol. From there, we can be certain of our low-cost results.
A very interesting point - I hadn’t thought about that, but now that I am, I’m wondering if we could expect HOB to be in the top 20 species (I think that’s what the affordable Plasmidsaurus gives) of an environmental soil sample. I’d expect them to be way down the list unless we were sampling them from somewhere that was deficient in other energy sources (like organics).
Unfortunately those organics (humic and fulvic acids, etc.) are likely to contaminate soil samples. My hope is that we can dilute these out sufficiently following an enrichment such that we don’t need the final 16,00 x g filtration step. But I think if you wanted to see how few of our final HOB were in the initial soil sample, you’d need at least the 15,100 x g SciSpin Micro Centrifuge, if not a second hand 16k+ model. And possibly a more expensive 16s service.
I’d like to think that we could get all of this with a UK service if we were offering sufficient samples. I think the key thing with Plasmidsaurus is that they are happy to stack individual samples from multiple customers, and have sufficient throughput to be able to do so.
They still haven’t responded to my enquiry. If they are doing the analysis of our samples in the UK then it may well be that they don’t have sufficient demand to offer all services at the advertised prices and turnarounds. In which case it becomes more than just the issue of international shipping of biological samples. In which case then yes, we’ll need to do our own PCR also.
I thought the AffiGEN kit was pretty much a cheaper alternative to the official Thermo version. As well as the cost advantage, Gemini suggests it’s more convenient in general use as you don’t need serial dilutions for 2 point calibrations. You would however still need serial dilutions for the 6 point validation assay.
Probably best checking this with DIYNAFLUOR. And IO Rodeo, if also trialing the Open Colourimeter Plus. You could produce an excellent paper comparing the 3 reagent sets and two hardware options - better still if you could get a local (or open source proponent) academic with a Qubit involved so you can also compare the industry standard hardware.