Difference between revisions of "Team:UMBC-Maryland/Experiments"

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<h2>Experiments &amp; Protocols</h2>
 
<h2>Experiments &amp; Protocols</h2>
  
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<h5>1% Agarose Gel</h5>
+
<h5>Agarose Gel</h5>
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<p>
+
 
Electrophoresis buffer contents:
 
Electrophoresis buffer contents:
 
<li>10 mL 50x TAE</li>
 
<li>10 mL 50x TAE</li>
 
<li>490 mL deionized water</li>
 
<li>490 mL deionized water</li>
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<li>50 μL Bullseye DNA SafeStain</li>
 
 
<br>  
 
<br>  
−
Contents of 1% Agarose gel:
+
Contents of Agarose gel:
−
<li>1g Agarose</li>
+
<li>1g Agarose (1%) or 2g Agarose (2%)</li>
 
<li>100 mL Electrophoresis buffer</li>
 
<li>100 mL Electrophoresis buffer</li>
 
<br>
 
<br>
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<li>When the solution is sufficiently cool, add 6 µL of EtBr.</li>
 
<li>When the solution is sufficiently cool, add 6 µL of EtBr.</li>
 
<li>Pour the solution into a bed with a comb levely placed within. Clear the bubbles from the surface and wait for the gel to polymerize.</li>
 
<li>Pour the solution into a bed with a comb levely placed within. Clear the bubbles from the surface and wait for the gel to polymerize.</li>
−
<li>Mix loading dye and sample in a 1:5 ratio and insert samples into wells. Place 10 µL of DNA ladder into the first well.</li> </p>
+
<li>Mix loading dye and sample in a 1:5 ratio and insert samples into wells. Place 10 µL of DNA ladder into the first well.</li>
 +
<br></br>
 +
<h5>Plasmid Extraction</h5>
 +
We used <a href="http://www.mn-net.com/ProductsBioanalysis/DNAandRNApurification/PlasmidDNApurificationeasyfastreliable/NucleoSpinPlasmidplasmidMiniprepkit/tabid/1379/language/en-US/Default.aspx">NucleoSpin® Plasmid Miniprep kit, Macherey-Nagel </a> according to the manufacturer's instructions.
 +
<br></br>
 +
<h5>Restriction Digest</h5>
 +
<li>8 μL 2.1 Buffer</li>
 +
<li>5 μL DNA</li>
 +
<li>2 μL EcoR1</li>
 +
<li>2 μL Pst1</li>
 +
<li>Incubate at 37oc for 1 hour</li>
 +
 
  
 
<h5>What should this page contain?</h5>
 
<h5>What should this page contain?</h5>

Revision as of 17:36, 14 August 2015

Team UMBC-Maryland banner.jpg



Experiments & Protocols

Agarose Gel
Electrophoresis buffer contents:
  • 10 mL 50x TAE
  • 490 mL deionized water

    • Contents of Agarose gel:
  • 1g Agarose (1%) or 2g Agarose (2%)
  • 100 mL Electrophoresis buffer

  • Mix agarose and electrophoresis buffer and microwave for 1.5 minutes.
  • When the solution is sufficiently cool, add 6 µL of EtBr.
  • Pour the solution into a bed with a comb levely placed within. Clear the bubbles from the surface and wait for the gel to polymerize.
  • Mix loading dye and sample in a 1:5 ratio and insert samples into wells. Place 10 µL of DNA ladder into the first well.


    • Plasmid Extraction
    We used NucleoSpin® Plasmid Miniprep kit, Macherey-Nagel according to the manufacturer's instructions.

    Restriction Digest
  • 8 μL 2.1 Buffer
  • 5 μL DNA
  • 2 μL EcoR1
  • 2 μL Pst1
  • Incubate at 37oc for 1 hour
    • What should this page contain?
    • Protocols
    • Experiments
    • Documentation of the development of your project

    Inspiration