Difference between revisions of "Team:Carnegie Mellon/Protocols"

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     $("#flip_miniprep").click(function(){
 
     $("#flip_miniprep").click(function(){
 
         $("#miniprep_p").slideToggle("slow");
 
         $("#miniprep_p").slideToggle("slow");
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    });
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});
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$(document).ready(function(){
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    $("#flip_restriction_enzyme_digest").click(function(){
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        $("#restriction_enzyme_digest_p").slideToggle("slow");
 
     });
 
     });
 
});
 
});
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     color: black;
 
     color: black;
 
}
 
}
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#restriction_enzyme_digest_p, #flip_restriction_enzyme_digest {
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    padding: 5px;
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    text-align: center;
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    border: solid 3px #c3c3c3;
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    background-color: #40e6ee; /* #69c94f; */
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    color: white;
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    width: 80%;
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    margin: auto;
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}
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#restriction_enzyme_digest_p {
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    padding: 30px;
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    display: none;
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    background-color: transparent;
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    width: 76%;
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    margin-left: auto;
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    text-align: left;
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    color: black;
 
}
 
}
 
/* /toggle menu code */
 
/* /toggle menu code */
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</ol>
 
</ol>
 
</div>
 
</div>
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<!-- __________________________ Restriction Enzyme Digest Protocol _____________ -->
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<div id = "flip_restriction_enzyme_digest">Restriction Enzyme Digest Protocol</div>
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<div id = "restriction_enzyme_digest_p">Restriction Enzyme Digest Protocol</div>
  
 
</body>
 
</body>
  
 
</html>
 
</html>

Revision as of 15:20, 10 August 2015

Under Construction.

This is almost as well-documented as Kim Kardashian's wedding.

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His-Tag Soluble Protein Extraction
  1. Centrifuge 1.5 ml of culture for 1min full speed at 15000 rpm.
  2. Remove supernatant.
  3. Add 250 µl of extraction buffer (1% octyl-beta-thioglucoside in 10mM Tris-Cl, pH7.5).
  4. Incubate at room temperature for 10 min.
Wash Beads
  1. Remove 50 µl of Ni-NTA in 10 mM Trischloride bead solution and place in new tube for each purification.
  2. Add 1 mL buffer (see table).
  3. Centrifuge at 800 rpm for 30 seconds.
  4. Remove supernatant.
  5. Repeat 3 times and resuspend in 1 mL of Bead Wash Solution.
Estrogen Sensor
Estrogen Sensor Protocol
MiniPrep Protocol
Minipreps of MACH Cells Expressing Flourescence
Purpose: To isolate plasmid DNA from MACH cells.

Procedure:

  1. Set up overnight cultures for miniprep.
  2. Make the following reaction recipe:
    • 5 mL LB
    • 5 µL Chlorophenical
    • 1 µL overnight colony
  3. Incubate in 37°C for 16-18 hours.
  4. Follow the Life Technologies Miniprep Kit Protocol.
Restriction Enzyme Digest Protocol
Restriction Enzyme Digest Protocol