Is Qubit good for multiple samples?

[seqtechno1]

Hello,

We've been having some quantitation issues recently, I'm wondering whether the Qubit fluorometers are good for processing lots of samples (96 at a time?)

[r.rosati]

Hi,
they should be... Once you've passed the 2-minute minimum incubation time, then the tubes are supposed to be stable for 3 hours (Qubit dsDNA HS Assay guide, page 2), although personally it never happened that I had them them sit for more than half an hour prior to reading them, so I can't speak from personal experience.
What kind of issues are you experiencing? Do you notice readings drop, or raise during time? Or do the readings just fluctuate? How do the calibrators perform at the end of the readings?
Or is your issue that at the end of the experiment, the proportion of libraries is uneven? In this case, besides quantity, are they even in quality?

[microgirl123]

What are you using for quantification now? I find the Qubit to be time-consuming for 96 samples - instead I use the Qubit reagents, white strip tubes, and a plate reader that can handle tubes.

[kmcarr]

What are you using for quantification now? I find the Qubit to be time-consuming for 96 samples - instead I use the Qubit reagents, white strip tubes, and a plate reader that can handle tubes.

microgirl,

What plate reader are you using?

[microgirl123]

Well. . . it's actually an old Stratagene qPCR instrument that will allow you to do a single endpoint read so it's only used as a "plate" reader now.

I'm not sure how well Qubit reactions would work in a plate since there's not a good way to really vortex the samples, but I haven't experimented. I half the volume of the reaction in the strip tubes both to save reagent costs and to allow more room for mixing.

[ychang]

Hi，microgirl,

your idea of using a qPCR instrument as a Qubit reader was great! could you give more detail of the protocol? Thanks!

[Carcharodon]

Well. . . it's actually an old Stratagene qPCR instrument that will allow you to do a single endpoint read so it's only used as a "plate" reader now.

I'm not sure how well Qubit reactions would work in a plate since there's not a good way to really vortex the samples, but I haven't experimented. I half the volume of the reaction in the strip tubes both to save reagent costs and to allow more room for mixing.

I use the Biotium AccuClear system, and for that you simply mix the DNA with the dye/buffer mixture by pipetting and allow it to sit for ~ 5 mins. I would bet good money that the same approach would work with the Qubit reagents.