Three assay sizes are available. The volume of components in each kit are sufficient for 1000, 250 and 50 (evaluation size) assays respectively. Kit components are shown in the table below.Table 1: Ultra High Sensitivity Assay Kit Contents
|DeNovix dsDNA Ultra High Sensitivity Dye (400x)||0.5 mL||125 µL||25 µL|
|DeNovix dsDNA Ultra High Sensitivity Buffer||200 mL||50 mL||10 mL|
|DeNovix dsDNA B Ultra High Sensitivity Enhancer (100x)||2 x 1 mL||0.5 mL||100 µL|
|300 pg/µL dsDNA Standard (calf thymus)||2 mL||1 mL||0.5 mL|
|0 pg/µL dsDNA Standard||2 mL||1 mL||0.5 mL|
Dynamic Range Considerations
The DeNovix Ultra High Sensitivity Assay enables dsDNA quantitation through the full dynamic range on Qubit® Fluorometers. However, the sensitivity of these instruments is inadequate to measure dsDNA at the low end of the assay range when following the standard protocol.
In order to measure dsDNA stock concentrations less than 10 pg/µL, sample volumes of 20 µL should be added to 190 µL working solution. For dsDNA concentrations above 10 pg/µL, the standard assay protocol is appropriate.
Installing DeNovix Assays on a Qubit® Fluorometer
For a Qubit® 3.0 / 4 Fluorometer:
Once the assay file is uploaded to the Qubit® Fluorometer, simply select the DeNovix High Sensitivity Assay to measure dsDNA samples.
Pay careful attention to pipetting accuracy when quantitating low picogram amounts of dsDNA.
Three replicates of dsDNA samples were measured on a Qubit® 3.0, a Qubit® 4 and a DeNovix QFX Fluorometer using the DeNovix Ultra High Sensitivity Assay. Samples above 10 pg/µL were prepared with 200 µL working solution and 10 µL sample. Samples below 10 pg/µL were prepared with190 µL working solution and 20 µL stock added. The data is presented in graphical and tabular form across the assay dynamic range below.
|Expected dsDNA||Qubit® 3.0||Qubit® 4||QFX|
|1||Measured Out of Range||0.06||0.006||1.16||0.31|
|0.5||Measured Out of Range||Measured Out of Range||0.25||1.19|
The DeNovix dsDNA Ultra High Sensitivity Assay enables specific, highly sensitive dsDNA quantitation across a wide dynamic range. The assay is available for use on a Qubit® Fluorometer through the simple application of downloadable assay files.
Qubit® is a registered trademark of Thermo Fisher Scientific and its subsidiaries.
Revised 19 Oct 2020