May 12, 2026
Nanodrop - dsDNA Quantification
- Owen Butler1,
- Raven Coffer1
- 1DAMP Lab
- DAMP Lab
Protocol Citation: Owen Butler, Raven Coffer 2026. Nanodrop - dsDNA Quantification. protocols.io https://dx.doi.org/10.17504/protocols.io.36wgqx1n5lk5/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: March 30, 2026
Last Modified: May 12, 2026
Protocol Integer ID: 314090
Keywords: dsdna quantification, dsdna quantification the goal, amount of dna, dna, absorbance ratio, purity, quantification
Abstract
The goal of this protocol is to quantify the amount of DNA in a sample, and determine the purity based on absorbance ratios.
Materials
**Nanodrop Materials**
- Pipette / tips
- Sample
- Blank
- KimWipe
- DI water
Quantifying dsDNA Sample Concentration and Purity
Nanodrop
Login to the computer, launch NanoEight Software.
Select dsDNA for double stranded DNA (this is the most common sample type for our workflows).
Figure 1. NanoDrop Eight software home page
To clean the pedestals, gently wipe the pedestals with some DI water on a KimWipe.
Note
Ensure there is no water left behind from cleaning.
Click "Ready" to continue.
Select the desired pedestals for blanking (up to eight pedestals depending on the number of pedestals you will require).
Figure 2. Selecting pedestals in the software
Add 1 µL of NFW or 1 µL Elution Buffer (depending on what your sample is in) to the center of the selected pedestals. Bubbles or off-centered liquids may disrupt and distort the read.
Close the arm and hit run to initiate blanking.
Figure 3. Running blank reads
Ensure the blanks were accepted. Repeat the cleaning process by gently wiping the pedestals with some DI water on a KimWipe.
Add 1 µL of sample to the center of the pedestal. Bubbles or off-centered liquids may disrupt and distort the read. Record the order of samples in your ELN. Click the play button at the top right to measure up to 8 samples.
Figure 4. Measuring samples
If you have more than 8 samples to read, wipe the pedestals down with DI water on a KimWipe.
Select the next column, load, and read your next samples. Repeat for however many samples you need to read, cleaning the pedestals in between each read of 8 samples.
Note
Do not perform another blank measurement between columns. All samples will have the same initial blank applied to their calculations.
End the experiment using the button in the top-right corner. Select the desired file format(s) (e.g., .csv) and export the data to Excel. Upload the exported files to your preferred storage location, then rename the samples to reflect their corresponding identities.
Figure 5. Ending run and exporting data
Repeat the cleaning process once more, lower the pedestal arms, and close the software.
Acknowledgements
Initially drafted by Raven Coffer. Uploaded by Zohkira Mukhammadyunusova. Edited by Owen Butler, Lena Landaverde, Kristen Sheldon, Molly Brennan.