NanoDrop Spectrophotometer Use

1. The NanoDrop machine is in the Cell lab at the back of the room.
   If the computer is off, turn it on and log in. Open NanoDrop software; link is on desktop.
   Choose the type of measurement you’d like to make* (Nucleic Acid, UV-Vis, Cell cultures, Protein, etc.)

2. A window opens asking you to load a water sample to initiate the machine: add 1.5 ul of millipore-filtered water to pedestal and lower the arm. Click "proceed". 
   When finished, clean top and bottom measurement pedestal surfaces with a Kimwipe tissue. 

3. Load 1.5 uL water or other appropriate blank (eg.TE buffer) on the pedestal and lower the top arm. Click on the “BLANK” button to run the blank measurement.
   When finished, clean the pedestal (top and bottom) with a Kimwipe.

4. Load a 1.5 uL of sample on the lower pedestal, lower the arm. If desired, enter the sample name in the window provided.
   Click the “Measure” button  – the display will then show the spectrum and show the calculated concentration. 
   Optional: When finished, click “show report”. From here you can save or print the report. The data can be saved to your own folder, P: drive, or disc.
   

5. Clean the pedestal and leave the arm down. Exit software.
   
   *Notes: If measuring protein samples, please see manual for more stringent cleaning procedures.
   If you are using “Nucleic Acid” you must select the extinction coefficient for the type of nucleic acid you are measuring in the upper right corner of the 
   nucleic acid page – RNA 40, DNA (double-stranded) 50, DNA (single-stranded) 33- so that the concentration will be correct for your sample
   Specifications: NanoDrop will measure dsDNA from 2 ng/uL to 3700 ng/uL without dilution; to 3000 ng/uL RNA; to 2400 ng/uL ssDNA.
   
   
   

F. L. Erickson, 1/2007