Team:TU Darmstadt/Protocols/DNA Quantification / NanoDrop

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Contents

DNA Quantification

Visual Comparison

It is possible to determine DNA concentrations by comparing the relative intensity of PCR bands and marker in agarose gel.

Nanodrop

DNA absorbs light at 260 nm and thus the concentration can be measured photometrically using the NanoDrop 1000 Spectrophotometer.

NanoDrop 1000

  1. Start the programme for the NanoDrop 1000 and click Nucleic Acids
  2. Pipette 1.5 µL of a water sample onto the lower measurement and click “OK”
  3. Load your blank and click Blank
  4. Load your DNA samples and click Measure. A 260/280 ration of 1.8 or above means the sample can be seen as pure
  5. At the end load your blank and click Measure to test the blank.
  6. If the measurement deviates more than 5 ng/µL from 0.0 ng/µL, load the blank again, click Blank and measure your DNA samples.

Shimadzu BioSpec Nano

Switch the NanoDrop ON make sure a wiper is attatched.

  1. Press F3 to blank with 2µl of dH2O eg.
  2. Put 1-2µl of solution of the NanoDrop lense
  3. Press F4 to measure
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