Team:Aberdeen Scotland/Project/Assay

From 2014.igem.org

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<p>What we aim to to with our assay is to reduce all to that the a small portable system. Meet the Diagnostics Backpack:</p>
<p>What we aim to to with our assay is to reduce all to that the a small portable system. Meet the Diagnostics Backpack:</p>
<img src="https://static.igem.org/mediawiki/2014/5/5c/Test_backpack.jpg" alt="Diagnostics Backpack">
<img src="https://static.igem.org/mediawiki/2014/5/5c/Test_backpack.jpg" alt="Diagnostics Backpack">
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<p>Protocol:</p>
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<ol>
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<li>Blood samples are taken from patients suspected to be suffering from Human African Sleeping Sickness (Trypanosomiasis), a blood parasite derived from the bite of an infected Tsetse fly. Blood collected is mixed with untransformed E. coli, this removes non-specific binding. Put blood and non-mimotope E. coli in tube together put on rocker for ~30mins.</li>
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<li>Suspension is passed through 0.22μm PTFE filter to remove cells and non-specific antibodies.</li>
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<li>Serum antibodies are bound to a Poly-L-Lysine-coated surface (premade eppendorffs), and incubated for ~2hours, rocking.</li>
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<li>Remove unbound serum and wash 3x PBS</li>
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<li>The surface is blocked to remove regions of non-specific binding with 3% w/v (pre-boiled & filtered) milk solution or 3% milk powder, incubated for ~1hour, rocking.</li>
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<li>Wash 1x PBS.</li>
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<li>Add prewashed LiTat1.3-(S) culture [see reagents], incubate for ~1 hour, rocking.</li>
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<li>Wash 3x PBS then add LiTat1.5-(R) culture, incubate for max ~1 hour, rocking.</li>
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<li>Wash 3x PBS then add medium.</li>
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</ol>
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Revision as of 23:27, 14 August 2014

Team:Aberdeen Scotland/Project/Assay - 2014.ogem.org



Assay

Comparing our assay to current ones


A typical assay as employed in the field currently looks something like this:

What we aim to to with our assay is to reduce all to that the a small portable system. Meet the Diagnostics Backpack:

Diagnostics Backpack

Protocol:

  1. Blood samples are taken from patients suspected to be suffering from Human African Sleeping Sickness (Trypanosomiasis), a blood parasite derived from the bite of an infected Tsetse fly. Blood collected is mixed with untransformed E. coli, this removes non-specific binding. Put blood and non-mimotope E. coli in tube together put on rocker for ~30mins.
  2. Suspension is passed through 0.22μm PTFE filter to remove cells and non-specific antibodies.
  3. Serum antibodies are bound to a Poly-L-Lysine-coated surface (premade eppendorffs), and incubated for ~2hours, rocking.
  4. Remove unbound serum and wash 3x PBS
  5. The surface is blocked to remove regions of non-specific binding with 3% w/v (pre-boiled & filtered) milk solution or 3% milk powder, incubated for ~1hour, rocking.
  6. Wash 1x PBS.
  7. Add prewashed LiTat1.3-(S) culture [see reagents], incubate for ~1 hour, rocking.
  8. Wash 3x PBS then add LiTat1.5-(R) culture, incubate for max ~1 hour, rocking.
  9. Wash 3x PBS then add medium.


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