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Team:EPF-Lausanne/Protocols/Gel purification
From 2011.igem.org
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{{:Team:EPF-Lausanne/Templates/Header|title=Gel purification}} | {{:Team:EPF-Lausanne/Templates/Header|title=Gel purification}} | ||
| - | '''Purpose''': | + | '''Purpose''': separate and extract products of a PCR |
| - | + | The PCR products are separated by gel electrophoresis, then mechanically cut out with the gel. The DNA is then extracted from each gel fragment. | |
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| + | Run a first electrophoresis with a low volume of PCR product, to verify the expected fragments were produced. If that is the case, run a new electrophoresis with all the PCR product, using a thicker combs to make larger wells. The well from our thick comb contain up to 25 or 30 ul. | ||
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| + | == Cutting the gel == | ||
First, cut out the band from the gel | First, cut out the band from the gel | ||
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* Hold the anti-UV shield with one hand and switch on the UV light | * Hold the anti-UV shield with one hand and switch on the UV light | ||
* Cut around the band, switch off the UV light and put the removed band in an eppendorf tube | * Cut around the band, switch off the UV light and put the removed band in an eppendorf tube | ||
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| + | == DNA Extraction == | ||
Then, to extract the DNA from the gel, use the Qiagen gel purification kit. | Then, to extract the DNA from the gel, use the Qiagen gel purification kit. | ||
Revision as of 07:35, 13 July 2011
Gel purification
Purpose: separate and extract products of a PCR
The PCR products are separated by gel electrophoresis, then mechanically cut out with the gel. The DNA is then extracted from each gel fragment.
Run a first electrophoresis with a low volume of PCR product, to verify the expected fragments were produced. If that is the case, run a new electrophoresis with all the PCR product, using a thicker combs to make larger wells. The well from our thick comb contain up to 25 or 30 ul.
Cutting the gel
First, cut out the band from the gel
- Take the black box off the UV plate
- Hold the anti-UV shield with one hand and switch on the UV light
- Cut around the band, switch off the UV light and put the removed band in an eppendorf tube
DNA Extraction
Then, to extract the DNA from the gel, use the Qiagen gel purification kit.
- To weight your samples, tare the scales with an empty tube
- In the elution step, when you have to centrifuge the column in a new Eppendorf tupe, make sure the lid of the tube flies behind in the centrifugator. It turns counterclockwise.
