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 Broad Range Dye (100x) | 2 x 1 mL | 0.5 mL | 100 µL |
| DeNovix dsDNA Broad Range Buffer | 250 mL | 50 mL | 10 mL |
| DeNovix dsDNA Broad Range Enhancer (100x) | 2 x 1 mL | 0.5 mL | 100 µL |
| 200 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
- Equilibrate all solutions to room temperature before use. Vortex, then centrifuge vials briefly to minimize reagent loss on the cap.
- Prepare working solution by mixing the dye and enhancer each with the assay buffer in a 1:100 ratio, e.g. 100 uL dye and 100 uL enhancer into 10 mL buffer. Scale volumes as needed to make enough volume to aliquot 190 µL of the mixture for each standard and unknown.
- For each standard or unknown sample, add 190 µL of the working solution to a labeled tube. Adjust volume when adding more or less than 10 µL of the unknown sample.
- Use thin-walled, clear UV-transparent 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 and 200 ng/µL standards and 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 |
|---|---|
| 0.2 – 200 ng/µL | 10 µL |
| 0.1 – 2 ng/µL | 20 µL |
| 200 – 1000 ng/µL | 2 µL |
| 1000 – 4000 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 Broad Range 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, lower the lid and tap Measure.
- Insert the 200 ng/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.
Reagent Storage
| Component | Protect from Light | Temperature |
|---|---|---|
| DeNovix dsDNA Broad Range Dye (100x) | Yes | 4°C - Room Temperature |
| DeNovix dsDNA Broad Range Buffer | Optional | 4°C - Room Temperature |
| DeNovix dsDNA Broad Range Enhancer (100x) | Optional | 4°C - Room Temperature |
| dsDNA Standards | Yes | 4°C |
01-MAR-2023
