Kit Contents
Kits are available in 1000, 250 and 50 (evaluation size) assays and include the components in Table 1. Safety data sheets are available at denovix.com/sds.
| Component | 1000 | 250 | EVAL |
|---|---|---|---|
| DeNovix dsDNA High Sensitivity Dye (100x) | 2 x 1 mL | 0.5 mL | 100 µL |
| DeNovix dsDNA High Sensitivity Buffer | 200 mL | 50 mL | 10 mL |
| 25 ng/µL dsDNA Standard (calf thymus) | 2 mL | 1 mL | 0.5 mL |
| 0 ng/µL dsDNA Standard | 2 mL | 1 mL | 0.5 mL |
Best Practices
Sample Prep
- 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 the dye with the assay buffer in a 1:100 ratio, e.g. 100 uL dye into 10 mL buffer. Scale volumes as needed to make enough volume to aliquot 190 µL of the mixture for each standard and unknown. Discard after 24 hours.
- For each standard or unknown sample, add 190 µL of the working solution into a labeled tube. Adjust volume when adding more or less than 10 µL of the unknown sample.
- Use thin-walled, clear 0.5 mL PCR tubes for assay measurements (DeNovix cat #TUBE-PCR-0.5-500 or equivalent).
- Add 10 µL of the 0 ng/µL, 25 ng/µL standards or 1 – 20 µL of 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. Total assay volume should remain 200 µL. Adjust working solution volume accordingly.
| Initial Sample Concentration | Recommended Sample Volume |
|---|---|
| 10 pg/µL – 25 ng/µL | 10 µL |
| 5 – 100 pg/µL | 20 µL |
| 25 – 125 ng/µL | 2 µL |
| 125 – 250 ng/µL | 1 µL |
Sample Measurement
- Launch the Fluoro dsDNA app using a DeNovix Fluorometer.
- Use the drop-down menu to select the correct LED source for the DeNovix dsDNA High Sensitivity Assay.
- Select the preferred standard curve method (2 point standards supplied) and then choose Generate New Standard Curve.
- Insert the 0 ng/µL dsDNA standard tube, lower the lid and tap the Measure button.
- Insert the 25 ng/µL dsDNA standard tube, lower the lid and tap the Measure button.
- After both standards are measured, tap the Samples button, insert a sample tube and tap Measure.
Reagent Storage
| Component | Protect from Light | Temperature |
|---|---|---|
| DeNovix dsDNA High Sensitivity Dye (100x) | Yes | 4°C - Room Temperature |
| DeNovix dsDNA High Sensitivity Buffer | Optional | 4°C - Room Temperature |
| dsDNA Standards | Yes | 4°C |
7-OCT-2024
