96 well fluorometer for genomic DNA quant?

[Lisa Jane Cross]

Hi there, we will soon be launching a high-throughput NGS service. We currently do all our genomic DNA quant on the Qubit (as we have had problems with the picogreen/LC480 quant). The Qubit however, really won't be suitable when our throughput rockets up.
Any suggestions/suppliers of a 96-well fluorometer out there anywhere? I'm not having much luck finding one..they are all single tube
Thanks!

[Vinz]

There are probably more than you want to test:
Biotek Synergy
Tecan Infinite
BMG labtech
berthold
Google plate reader fluorescence for more...

[TonyBrooks]

If you have a qPCR machine capable of end-point reading (alellic discriminiation), it may be worth investigating whether you can read Picogreen fluoresence on it.
We have used our ABI 7900HT to read Picogreen plates, just set one marker to VIC and the other to SYBR with no (ROX) normalisation. The raw SYBR (I think) reads should be proportional to the mass of DNA in the sample. Just include a standard curve from the Lambda DNA, which is usually included in the Picogreen kit to determine the absolute amount.

[SarahNGS]

yes you want to look for a "fluorescence plate reader". PerkinElmer makes several including the EnSpire and Victor. They both work great for Pico Green.

[Ber7702]

For cheap and cheerful look at the Molecular Devices F3 or F5 Filtermax plate readers. For expensive and funky looking, you could take a look at the Trinean DropSense. It's basically a 96 well plate version of the Nanodrop.

[Old guy]

I have access to a Synergy HT and about 2,000 samples to quantify. Any suggestions on a protocol? Whose picogreen do you use and what is the most used standard? Thanks

[SarahNGS]

I have access to a Synergy HT and about 2,000 samples to quantify. Any suggestions on a protocol? Whose picogreen do you use and what is the most used standard? Thanks

Invitrogen makes a PicoGreen kit containing a standard: http://products.invitrogen.com/ivgn/...i8hAAAAA%3D%3D

Also Promega makes the QuantiFluor system:

http://www.promega.com/products/dna-and-rna-purification/dna-and-rna-quantitation/quantifluor-dsdna-system/

which also contains a lambda DNA standard. I have not used this one myself but the price is lower than the Invitrogen kit.

I've never used a synergy reader but basically you'd set up a standard curve then measure your fluorescence intensity against it. Depending on the reader, I've done either intensity vs. concentration as well as a log-log plot; use whichever gives you the straightest calibration curve. Ideally one does triplicate wells, but for 2000 samples that may be too much.