Team:Washington/NANODROP PROTOCOL

From 2013.igem.org

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Nanodrop Protocol

Procedure from: http://openwetware.org/wiki/Nanodrop

Typical DNA peak as measured by the Nanodrop on PC

1. Before starting the software module, clean the sample surfaces with DI water to remove any dried sample that might be present.
1. Alternatively, you can clean the sample surfaces with a Kimwipe moistened with 70% ethanol.
2. Open the Nanodrop program and the appropriate module (e.g., “Nucleic Acid”).
3. Wipe off the top and bottom sensors of the instrument with a Kimwipe. These are just the polished ends of fiber optic cable, so wiping is sufficient to prevent carryover.
4. Pipette 1-2 μL of DI water onto the sensor. Bring down the lever arm.
5. Follow the onscreen prompts to calibrate.
6. Wipe the sensors and pipette on 1-2 μL of the corresponding blank (Buffer EB or whatever solution your prep is in). Bring down the lever arm.
7. Follow the onscreen prompts to blank.
8. Wipe the sensors and pipette on 1-2 μL of your sample. Bring down the lever arm.
9. Click Measure and record the concentration measured.
10. For DNA, the peak should be at 260 nm, and as a general rule, the 260/280 ratio should be between 1.8 and 2.0.
11. To test multiple samples, just wipe the sensor in between measurements with a Kimwipe. Recalibration or re-blanking is not necessary.
12. Clean the sample surfaces once more after you are finished.