Team:Paris Saclay/ethanol

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= '''Ethanol precipitation''' =
= '''Ethanol precipitation''' =
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This method purifies and gathers DNA in a mixing
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This method is used to purify and/or concentrate DNA, RNA and polysaccarides.
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Add in a volume of DNA :
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1. 3 volumes of ethanol 100%
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'''Protocol :'''
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2. 1/10 of volume of sodium acetate 3M
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1. Measure the volume of the DNA sample.
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3. 1µL of glycogen
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2.     Add 3x volumes of ethanol 100%
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4. Let the solution at -20°C during 30min
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3. Add 1/10x volume of sodium acetate (3 M, pH 5.2)
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5. Centrifuge at maximal speed during 30min at 4°C
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4. Add 1µL of glycogen
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6. Put away the supernatant
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5. Incubate the solution at -20°C for 30min or more
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7. Wash with 1mL of ethanol 70%
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6. Centrifuge at > 14,000 x g for 30min at 4°C
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8. Centrifuge 10-15 min at 4°C
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7. Discard supernatant being careful not to throw out DNA pellet which may or may not be visible.
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9. Put away the supernatant, dry (speed vacuum 4 min)
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8. Wash with 1mL of ethanol 70%
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10. Repeat with water or TE 
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9. Centrifuge at > 14,000 x g for 15min at 4°C
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10. Discard supernatant, air dry pellet and dissolve pellet in desired buffer or water

Latest revision as of 14:23, 4 October 2013

Navigation Home Team Project Labwork Notebook Human practices

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



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