Difference between revisions of "Team:Calgary/Experiments/imposter"

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<h1>Our Experiments</h1>
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<h2>Rehydration of Registry DNA</h2>
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<table border="0">
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<tr><td>
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<p><b>Experimental Details and Rationale</b></p>
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</td>
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<td>
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<p>Registry DNA was rehydrated for completion of the Interlab Study. Also, Part BBa_K934001 (phaC1-A-B1) was rehydrated by the Secretion subgroup and transformed into our chassis so that P(3HB) was produced and preliminary secretion assays could be performed before the Synthesis subgroup had completed their cloning.</p>
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</tr></td>
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<tr><td>
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<p><b>Materials</b></p>
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</td>
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<td>
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<p>iGEM 2017 distribution kit</p>
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<p>ddH₂O</p>
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</tr></td>
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<tr><td>
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<p><b>Protocol</b></p>
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</td>
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<td>
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<ol><p>
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<li>Add 10 μL of ddH₂O to the desired well.</li>
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<li>Pipette up and down 3-5 times.</li>
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<li>Incubate at room temperature for 10 minutes.</li>
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<li>Transform cells with 1 μL of rehydrated DNA. Store the remaining amount at -20°C.</li>
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</p></ol>
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</tr></td>
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</table>
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<h2>Rehydration of IDT Synthesized DNA</h2>
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<table border="0">
 +
 +
<tr><td>
 +
<p><b>Experimental Details and Rationale</b></p>
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</td>
 +
<td>
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<p>Our genetic parts were ordered from IDT and arrived as a dry, lyophilized powder. They were resuspended in aqueous solution for cloning into pSB1c3 or pET29B vectors and to ligate multiple parts together. </p>
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</tr></td>
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<tr><td>
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<p><b>Materials</b></p>
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</td>
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<td>
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<p>Synthesized DNA from IDT (gBlocks)</p>
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<p>ddH₂O</p>
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</tr></td>
 +
 +
<tr><td>
 +
<p><b>Protocol</b></p>
 +
</td>
 +
<td>
 +
<ol><p>
 +
<li>Centrifuge tube containing the synthesized DNA for 3-5 seconds at 3000g to ensure that all material is at the bottom of the tube.</li>
 +
<li>Add ddH₂O to reach a final concentration of 50 ng/μL.</li>
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<li>Vortex.</li>
 +
<li>Incubate at 50°C for 20 minutes.</li>
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<li>Briefly vortex and centrifuge . Store at -80°C.</li>
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</p></ol>
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</tr></td>
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</table>
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</html>
 
</html>

Revision as of 22:57, 18 July 2017

Header

Our Experiments

Rehydration of Registry DNA

Experimental Details and Rationale

Registry DNA was rehydrated for completion of the Interlab Study. Also, Part BBa_K934001 (phaC1-A-B1) was rehydrated by the Secretion subgroup and transformed into our chassis so that P(3HB) was produced and preliminary secretion assays could be performed before the Synthesis subgroup had completed their cloning.

Materials

iGEM 2017 distribution kit

ddH₂O

Protocol

  1. Add 10 μL of ddH₂O to the desired well.
  2. Pipette up and down 3-5 times.
  3. Incubate at room temperature for 10 minutes.
  4. Transform cells with 1 μL of rehydrated DNA. Store the remaining amount at -20°C.

Rehydration of IDT Synthesized DNA

Experimental Details and Rationale

Our genetic parts were ordered from IDT and arrived as a dry, lyophilized powder. They were resuspended in aqueous solution for cloning into pSB1c3 or pET29B vectors and to ligate multiple parts together.

Materials

Synthesized DNA from IDT (gBlocks)

ddH₂O

Protocol

  1. Centrifuge tube containing the synthesized DNA for 3-5 seconds at 3000g to ensure that all material is at the bottom of the tube.
  2. Add ddH₂O to reach a final concentration of 50 ng/μL.
  3. Vortex.
  4. Incubate at 50°C for 20 minutes.
  5. Briefly vortex and centrifuge . Store at -80°C.

References

Rose, C., Parker, A., Jefferson, B., & Cartmell, E. (2015). The Characterization of Feces and Urine: A Review of the Literature to Inform Advanced Treatment Technology. Critical Reviews In Environmental Science And Technology, 45(17), 1827-1879. http://dx.doi.org/10.1080/10643389.2014.1000761