Team:Newcastle/Project/Labwork/OurProtocols/EthanolPrecip
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+ | =Ethanol Precipitation= | ||
+ | <font color="green">'''''This protocol is designed to remove the ethanol which is likely to be present in a DNA sample once it has been midi-prepped (the ethanol is found in Wash Solution 2).'''''</font> | ||
+ | |||
+ | ===What you will need=== | ||
+ | * 3M Sodium Acetate (pH 5.2) | ||
+ | * 100% ethanol | ||
+ | * 70% ethanol | ||
+ | * Deionised water or buffer (10mM) | ||
+ | * Ice bucket (with ice) | ||
+ | * Eppendorf Tubes | ||
+ | * Centrifuge | ||
+ | <br> | ||
+ | ===Procedure=== | ||
+ | # Measure the DNA sample volume and then calculate 1/10 of this volume. This is the amount of sodium acetate that needs to be added to the sample - pipette this volume of sodium acetate into the DNA sample. | ||
+ | <br> | ||
+ | <font color="blue">''For example if the DNA sample measures 2ml calculate 10% of this volume (which is 0.2ml). Take up 0.2ml of the sodium acetate and add it to the DNA sample - this should make up a total volume of 2.2ml.''</font> | ||
+ | <br> | ||
+ | # Add 100% ethanol to the solution. To work out the amount of 100% ethanol needed, multiply the volume of the DNA sample (plus sodium acetate) by 3. | ||
+ | <br> | ||
+ | <font color="blue">''For example if the DNA sample (plus sodium acetate) makes up a volume of 2.2ml, multiply the 2.2ml by 3, which gives the amount of 6.6ml. Add this volume of ethanol to the DNA (plus sodium acetate) solution, which should now give a total volume of 8.8ml.''</font> | ||
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Revision as of 19:42, 9 August 2009
Ethanol Precipitation
This protocol is designed to remove the ethanol which is likely to be present in a DNA sample once it has been midi-prepped (the ethanol is found in Wash Solution 2).
What you will need
- 3M Sodium Acetate (pH 5.2)
- 100% ethanol
- 70% ethanol
- Deionised water or buffer (10mM)
- Ice bucket (with ice)
- Eppendorf Tubes
- Centrifuge
Procedure
- Measure the DNA sample volume and then calculate 1/10 of this volume. This is the amount of sodium acetate that needs to be added to the sample - pipette this volume of sodium acetate into the DNA sample.
For example if the DNA sample measures 2ml calculate 10% of this volume (which is 0.2ml). Take up 0.2ml of the sodium acetate and add it to the DNA sample - this should make up a total volume of 2.2ml.
- Add 100% ethanol to the solution. To work out the amount of 100% ethanol needed, multiply the volume of the DNA sample (plus sodium acetate) by 3.
For example if the DNA sample (plus sodium acetate) makes up a volume of 2.2ml, multiply the 2.2ml by 3, which gives the amount of 6.6ml. Add this volume of ethanol to the DNA (plus sodium acetate) solution, which should now give a total volume of 8.8ml.
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