Troy.moench (Talk | contribs) |
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− | <p>The solution is syringe filtered through a 0.45 micrometer filter to trap bacteria bound to phage tails and allow any unbound phage tail through. This prevents false positives by ridding the solution of excess phage tail for | + | <p class="big">The solution is syringe filtered through a 0.45 micrometer filter to trap bacteria bound to phage tails and allow any unbound phage tail through. This prevents false positives by ridding the solution of excess phage tail for |
step three. | step three. | ||
</p> | </p> | ||
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<h2 class="alt">Step three</h2> | <h2 class="alt">Step three</h2> | ||
− | <p>Substrate is added to the filter which will be cleaved by the enzyme. Color change occurs if <i>E. coli</i> is present. Note, each <i>E. coli</i> cell can bind hundreds of phage tail and each tail may bind several enzyme, which allows for high sensitivity. </p> | + | <p class="big">Substrate is added to the filter which will be cleaved by the enzyme. Color change occurs if <i>E. coli</i> is present. Note, each <i>E. coli</i> cell can bind hundreds of phage tail and each tail may bind several enzyme, which allows for high sensitivity. </p> |
<img src="https://static.igem.org/mediawiki/2017/6/6d/T--WLC-Milwaukee--Filter_Paper.gif"> | <img src="https://static.igem.org/mediawiki/2017/6/6d/T--WLC-Milwaukee--Filter_Paper.gif"> |
Revision as of 04:53, 1 November 2017
Description
Step one
His-tag purified phage tail tip conjugated to a colorometric enzyme is added to a water sample by drawing water into a syringe. The tail tip recognizes LamB, an outer membrane protein on E. coli and binds to any E. coli present in the sample.
Step two
The solution is syringe filtered through a 0.45 micrometer filter to trap bacteria bound to phage tails and allow any unbound phage tail through. This prevents false positives by ridding the solution of excess phage tail for step three.
Step three
Substrate is added to the filter which will be cleaved by the enzyme. Color change occurs if E. coli is present. Note, each E. coli cell can bind hundreds of phage tail and each tail may bind several enzyme, which allows for high sensitivity.