Difference between revisions of "Elute the released products"
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(Created page with ''''The Procedure''' #Remove the DNA fragments from the agarose gel with a clean scalpel. Try to get as small a piece of gel as possible, without touching the DNA. #Weigh the gel...') |
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#Weigh the gel slice in a clorless tube. Add 3 times the volume of Buffer QG to 1 volume of gel. | #Weigh the gel slice in a clorless tube. Add 3 times the volume of Buffer QG to 1 volume of gel. | ||
#Incubate at 50°C for 10 minutes, or until all the gel has dissolved. Mix the tube using the vortex mixer every 2 or 3 minutes to make sure it dissolves. | #Incubate at 50°C for 10 minutes, or until all the gel has dissolved. Mix the tube using the vortex mixer every 2 or 3 minutes to make sure it dissolves. | ||
| − | #Check if the mixture is yellow in color. If it is, it means that its pH is around 7.5. This is the pH at which the adsorbtion (stickiness)of the QIAquick membrane is most efficient. If it is orange or violet, it is too acidic. | + | #Check if the mixture is yellow in color. If it is, it means that its pH is around 7.5. This is the pH at which the adsorbtion (stickiness)of the QIAquick membrane is most efficient. If it is orange or violet, it is too acidic. Add 10µl of 3 M sodium acetate (pH 5.0) and mix, if that is the case. |
| + | #Add isopropanol of a volume equal to the volume of the gel to the sample and mix. The isopropanol precipitates the DNA. (?) | ||
Revision as of 15:19, 15 June 2009
The Procedure
- Remove the DNA fragments from the agarose gel with a clean scalpel. Try to get as small a piece of gel as possible, without touching the DNA.
- Weigh the gel slice in a clorless tube. Add 3 times the volume of Buffer QG to 1 volume of gel.
- Incubate at 50°C for 10 minutes, or until all the gel has dissolved. Mix the tube using the vortex mixer every 2 or 3 minutes to make sure it dissolves.
- Check if the mixture is yellow in color. If it is, it means that its pH is around 7.5. This is the pH at which the adsorbtion (stickiness)of the QIAquick membrane is most efficient. If it is orange or violet, it is too acidic. Add 10µl of 3 M sodium acetate (pH 5.0) and mix, if that is the case.
- Add isopropanol of a volume equal to the volume of the gel to the sample and mix. The isopropanol precipitates the DNA. (?)
