Digestion NotI
Add: - nuclease-free water qsp 50 µL > 47 µL - 5µL 10X Buffer O- 1µL DNA (1 µg/µL) - 1 µL NotI 1 (10U/µL) *. 
Mix gently and spin down for a few seconds.
Incubate at 37°C for 1-16 hours.
The digestion reaction may be scaled either up or down. 
It's possible to process extended incubation by adding 0.25U/µg of DNA in 50 µL of reaction volume.
incubation at 80°C for 20 min.
Measure the volume of the DNA sample > 50 µL
Add 1/10 volume of sodium acetate, pH 5.2, (final concentration of 0.3 M) 
- These amounts assume that the DNA is in TE only; if DNA is in a solution containing salt, adjust salt accordingly to achieve the correct final concentration. > 5 µL.
Mix well.
Add 2 to 2.5 volumes of cold 100% ethanol (calculated after salt addition).> 110 (2V).
Mix well.
Place on ice or at -20 degrees C for >20 minutes.
Spin a maximum speed in a microfuge 10-15 min.
Carefully decant supernatant.
Add 1 ml 70% ethanol.
Mix.
Spin briefly.
Carefully decant supernatant.
Air dry or briefly vacuum dry pellet.
Resuspend pellet in the appropriate volume of TE or water.
