Kits are available in 1000, 250 and 50 (evaluation size) assays and include the components in Table 1.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|
- Allow all solutions to equilibrate to room temperature before use. Vortex, then centrifuge vials briefly before opening to minimize reagent loss on the cap.
- Prepare working solution by mixing 10 mL of the assay buffer with 25 µL of the dye and 100 µL of the enhancer. Scale volumes as needed to make enough volume to aliquot 200 µL of the mixture for each standard and unknown.
- For each standard or unknown sample, add 200 µL of the working solution into a labeled tube.
- Use only thin-walled, clear 0.5 mL PCR tubes for assay measurements (DeNovix cat #TUBE-PCR-0.5-500 or equivalent). Label only the top, not the sides of an assay tube.
- Add 10 µL of the 0 pg/µL, 300 pg/µL standards or unknown DNA samples to the respective tubes and mix well.
- Incubate assay tubes at room temperature for 5 minutes.
Recommended Sample Volume
These recommendations ensure that sample concentrations are within the total mass detection limits of the assay.
|Initial Sample Concentration||Recommended Sample Volume|
|0.5 – 300 pg/µL||10 µL|
- Launch the Fluoro dsDNA app using a DeNovix Fluorometer.
- Use the drop-down menu to select the correct LED source for the DeNovix dsDNA Ultra High Sensitivity Assay.
- Select the preferred standard curve method (2 point standards supplied) and then choose Generate New Standard Curve.
- Insert the 0 pg/µL dsDNA standard, lower the lid and tap Measure.
- Insert the 300 pg/µL dsDNA standard, lower the lid and tap Measure.
- After both standards are measured, tap the Samples button, insert a sample tube and tap Measure.
|Component||Protect from Light||Temperature|
|DeNovix dsDNA Ultra High Sensitivity Dye (400x)||Yes||4°C - Room Temperature|
|DeNovix dsDNA Ultra High Sensitivity Buffer||Optional||4°C - Room Temperature|
|DeNovix dsDNA UltraHigh Sensitivity Enhancer (100x)||Optional||4°C - Room Temperature|
Revised 19 Oct 2021