Team:EPF-Lausanne/Protocol/DNAConcentration

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Revision as of 14:35, 13 July 2012

Protocol: DNA concentration measurement


  • Take a new 6 µl aliquote of the DNA (under the hood) and put back the main DNA tube in the fridge.
  • Go to the room by the E. coli lab (not on Friday morning!) with:
    • The 6 µl aliquote
    • A 10 µl pipete
    • Eventually some 5 µl of TE buffer (they migh have some).
  • The machine is the NanoDrop Spectrophotometer.
  • On the computer, click on "Nucleic Acid".
  • Add 2 µl of (nuclease free) water to the machine's tip as you are asked to and measure.
  • Clean tips (both sides) with a tissue.
  • Add 2 µl of TE buffer and click on "Blank".
  • Clean tips (both sides) with a tissue.
  • Add 2 µl of DNA and click "Measure".
  • Clean tips (both sides) with a tissue.
  • Take 2 measurements per sample and average.
  • Click on exit.

The important numbers are:

  • 260/280 ratio, must be > 1.8
  • 260/230 ratio, must be > 2 (too big, > 2.5? , might mean too much salts)
  • Of course the DNA concentration.