Team:CU-Boulder/Project/Kit/DNAPurification

From 2013.igem.org

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<dt>Homemade Miniprep System</dt>
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<dt>Gel Purification of DNA</dt>
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<dd> We developed a method for making our own miniprep spin columns that is much cheaper than commercial plasmid purification kits. Additionally, we tested and validated an effective method of recycling mini-columns to even further reduce cost and waste!</dd>
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<dd>This simple procedure utilizes filter paper and dialysis tubing to obtain concentrated DNA from an agarose gel, eliminating the need for commercial gel purification kits.</dd>
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<dd><b>GoPro Video</b></dd>
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<video width="640" height="360" controls>
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<source src="https://static.igem.org/mediawiki/2013/1/11/GelExtraction.webm" type="video/webm">
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<source src="https://static.igem.org/mediawiki/2013/b/be/GelExtraction.mp4" type="video/mp4">
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<li><a href="https://2013.igem.org/Team:CU-Boulder/Notebook/Protocols/DNAPurification">Step-By-Step Protocol</a></li>
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<dt>Homemade Miniprep Spin Column and Recycling System</dt>
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<p></p>
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<dd> We developed a method for making our own miniprep spin columns that is much cheaper than commercial plasmid purification kits. Additionally, we tested and validated an effective method of recycling mini-columns to even further reduce cost and waste!</dd>
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<p></p>
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<img src="https://static.igem.org/mediawiki/2013/5/56/Miniprep.jpeg" width = "250">
<img src="https://static.igem.org/mediawiki/2013/5/56/Miniprep.jpeg" width = "250">
<img src="https://static.igem.org/mediawiki/2013/6/6a/Recycleblowup.jpeg" width="700" >
<img src="https://static.igem.org/mediawiki/2013/6/6a/Recycleblowup.jpeg" width="700" >
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Columns were used to purify RFP (red) expressing plasmid. Columns were recycled as per the recycling protocol (left) or not (right) and then used to purify AmilCP (blue) expressing plasmid. The elutant was transformed and plated. Recycled columns resulted in 100% AmilCP (blue) producing colonies. Non-recycled columns produced ~ 50% AmilCP (blue) producing colonies and ~50% RFP producing colonies resulted from contaminating plasmid from the previous purification.  Recycling works.
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Columns were used to purify RFP (red) expressing plasmid. Columns were recycled as per the recycling protocol (left) or not (right) and then used to purify AmilCP (blue) expressing plasmid. The elutant was transformed and plated. Recycled columns resulted in 100% AmilCP (blue) producing colonies. Non-recycled columns produced ~50% AmilCP (blue) producing colonies and ~50% RFP producing colonies resulting from contaminating plasmid from the previous purification.  Recycling works!
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<dl>
 
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<dt>Gel Purification of DNA</dt>
 
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<p></p>
 
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<dd>This simple procedure utilizes filter paper and dialysis tubing to obtain concentrated DNA from an agarose gel, eliminating the need for commercial gel purification kits.</dd>
 
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<ul>
 
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<p>
 
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Video of filter paper+dialysis tubing method of DNA gel purification (500% speed)
 
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<video width="640" height="360" controls>
 
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<source src="https://static.igem.org/mediawiki/2013/d/dd/DNA500p.mp4" type="video/mp4">
 
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</video>
 
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<li><a href="https://2013.igem.org/Team:CU-Boulder/Notebook/Protocols/DNAPurification">Gel Purification of DNA</a></li>
 
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Latest revision as of 17:01, 27 October 2013

Gel Purification of DNA

This simple procedure utilizes filter paper and dialysis tubing to obtain concentrated DNA from an agarose gel, eliminating the need for commercial gel purification kits.
Homemade Miniprep Spin Column and Recycling System

We developed a method for making our own miniprep spin columns that is much cheaper than commercial plasmid purification kits. Additionally, we tested and validated an effective method of recycling mini-columns to even further reduce cost and waste!

Columns were used to purify RFP (red) expressing plasmid. Columns were recycled as per the recycling protocol (left) or not (right) and then used to purify AmilCP (blue) expressing plasmid. The elutant was transformed and plated. Recycled columns resulted in 100% AmilCP (blue) producing colonies. Non-recycled columns produced ~50% AmilCP (blue) producing colonies and ~50% RFP producing colonies resulting from contaminating plasmid from the previous purification. Recycling works!