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.