"
Team:UNIPV-Pavia/Protocols/Precipitation
From 2009.igem.org
(Difference between revisions)
(→DNA precipitation with sodium acetate) |
|||
| Line 38: | Line 38: | ||
'''Materials needed:''' | '''Materials needed:''' | ||
| - | *''' | + | *'''Sodium acetate''' |
| - | *''' | + | *'''Absolute ethanol''' |
| - | *''' | + | *'''Ethanol 70%''' |
*'''ddH2O''' | *'''ddH2O''' | ||
| - | *''' | + | *'''Previously cut plasmid''' |
<br> | <br> | ||
*We use it only if the miniprep did not give us a high yield of DNA | *We use it only if the miniprep did not give us a high yield of DNA | ||
Revision as of 15:01, 25 June 2009
|
DNA precipitation with sodium acetate
(estimated time: 1 hour)
Materials needed:
- Sodium acetate
- Absolute ethanol
- Ethanol 70%
- ddH2O
- Previously cut plasmid
- We use it only if the miniprep did not give us a high yield of DNA
- Add 1/10 digestion volume of sodium acetate 3 M
- Add 2.5 digestion volume of absolute ethanol
- Freeze at -80°C for 30 min
- Centrifuge at 13000 rpm, 4°C for 20 min
- Decant supernatant
- Add 50 µl of 70% ethanol
- Centrifuge at 13000 rpm, 4°C for 20 min
- Remove all supernatant with a pipette
- Air dry pellet until ethanol is totally removed
- Elute with 5 µl ddH2O
