Team:Paris Saclay/ethanol
From 2013.igem.org
(Difference between revisions)
(→Ethanol precipitation) |
(→Ethanol precipitation) |
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This method is used to purify and/or concentrate DNA, RNA and polysaccarides. | This method is used to purify and/or concentrate DNA, RNA and polysaccarides. | ||
- | + | '''Protocol :''' | |
- | 1. | + | 1. Measure the volume of the DNA sample. |
- | 2. | + | 2. Add 3x volumes of ethanol 100% |
- | 3. | + | 3. Add 1/10x volume of sodium acetate (3 M, pH 5.2) |
- | 4. | + | 4. Add 1µL of glycogen |
- | 5. | + | 5. Incubate the solution at -20°C for 30min or more |
- | 6. | + | 6. Centrifuge at > 14,000 x g for 30min at 4°C |
- | 7. | + | 7. Discard supernatant being careful not to throw out DNA pellet which may or may not be visible. |
- | 8. | + | 8. Wash with 1mL of ethanol 70% |
- | 9. | + | 9. Centrifuge at > 14,000 x g for 15min at 4°C |
- | + | ||
- | + | ||
+ | 10. Discard supernatant, air dry pellet and dissolve pellet in desired buffer or water | ||
Latest revision as of 14:23, 4 October 2013
Ethanol precipitation
This method is used to purify and/or concentrate DNA, RNA and polysaccarides.
Protocol :
1. Measure the volume of the DNA sample.
2. Add 3x volumes of ethanol 100%
3. Add 1/10x volume of sodium acetate (3 M, pH 5.2)
4. Add 1µL of glycogen
5. Incubate the solution at -20°C for 30min or more
6. Centrifuge at > 14,000 x g for 30min at 4°C
7. Discard supernatant being careful not to throw out DNA pellet which may or may not be visible.
8. Wash with 1mL of ethanol 70%
9. Centrifuge at > 14,000 x g for 15min at 4°C
10. Discard supernatant, air dry pellet and dissolve pellet in desired buffer or water