"
Finish MaxiPrep
Removal of remaining salts: Added 500ul of TE to the dried DNA pellet. Put in incubator for 1h30 to dissolve the pellet. Centrifuged at 10'000 rpm for 10min (in eppendorfs). Put the supernatent into another eppendorf and put in storage, in a yellow box in the cell lab fridge.
Quantifying the amount of DNA:
- RO plasmid: 407.2 ng/µl
- LovTAP plasmid: 294.5 ng/µl
For both the 260/280 ratio was 1.865 and 260/230 was 2.315,
Protocol: DNA Concentration Measurement
- Take a 6 µl aliquote of the DNA and put back the main DNA tube in the fridge.
- Go to the room by the E.Coli lab (LBTM, not on Friday morning!) with:
- The 6 µl aliquote
- A 10 µl pipet
- Optionally, the buffer you used for DNA elution (there might be some next to the machine).
- The machine is the NanoDrop Spectrophotometer.
- On the computer, click on "Nucleic Acid".
- Put a 2 µl drop of (nuclease-free) water on the machine's tip as you are asked to and measure.
- Clean tips (both sides) with a quarter of tissue.
- Add 2 µl of the buffer you use and click on "Blank".
- Clean tips (both sides).
- Add 2 µl of your DNA sample and click "Measure".
- Clean tips (both sides) with a tissue.
- Take 2 measurements per sample (for averaging).
- Print the report when you are done
- 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.
