Luckau Protocols:NanoDrop
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NanoDrop Protocol
Purpose
The NanoDrop is used to quantify genetic material. Although there are many different methods to quantifying DNA, the NanoDrop is a spectrophotometer that calculates absorbance of a sample across different wavelengths.
Protocol
- Assemble cafeteria tray
- Kim Wipes
- 2µL pipette
- pipette tips
- gloves
- USB drive
- water and buffer tubes
- samples
- The NanoDrop is located in North Life Sciences, room 325A
- Open the NanoDrop software, "ND1000" on the desktop
- Choose "Nucleic Acids"
- Initialize the instrument
- Place 2µL of NanoPure water on the pedestal
- Press OK
- when it's done, wipe pedestal and top connector with Kim Wipe
- Calibrate the instrument
- Place 2µL of elution buffer on the pedestal
- For Clark's lab, elution buffer is 10mM Tris-Cl, pH 8.4
- Click "Blank"
- when it's done, wipe pedestal and top connector with Kim Wipe
- Measure sample
- Place 2µL of sample on the pedestal
- Enter sample ID
- Click "Measure"
- wipe pedestal and top connector with Kim Wipe after each sample
- Re-calibrate the instrument each 10 samples, or so, by going back to step 6 and clicking "Re-Blank" instead of "Blank"
- Save data
- Click "Show Report"
- Click "Report", "Export Data"
- Click "Table Report"
- save to your USB drive, as "YYYYMMDD_NanoDrop", use "A" "B" "C", etc to designate multiple saved files in the same day
- Re-initialize the instrument each 30-50 samples, by going back to step 3
- remember to save your data before your re-initialize
- Clean up thoroughly! Other people use this instrument!
- don't forget to enter data into spreadsheet