Difference between revisions of "Team:Warwick/Protocols"
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<h4>Experiment 1: Testing the binding of specifically designed DNA strands to glass slides</h4> | <h4>Experiment 1: Testing the binding of specifically designed DNA strands to glass slides</h4> | ||
| − | <br>Glass slides were prepared <a href=" | + | <br>Glass slides were prepared <a href="https://static.igem.org/mediawiki/2015/f/f8/WarwickGlassSlideProtocol.pdf">Glass Slide Preperation Protocol</a> by being cleaned and functionalised (with HCl and GOPTS respectively). |
<br>Specifically designed oligonucleotides containing zinc finger binding domains were introduced to the slides. | <br>Specifically designed oligonucleotides containing zinc finger binding domains were introduced to the slides. | ||
<br>These oligonucleotides comprise of a general adaptor strand, a specific short strand and a specific long strand. | <br>These oligonucleotides comprise of a general adaptor strand, a specific short strand and a specific long strand. | ||
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<p> | <p> | ||
<H3> Experiment 3: Reciprocal experiment - binding of fluorescently labelled oligonucleotides to immobilised cells </H3> | <H3> Experiment 3: Reciprocal experiment - binding of fluorescently labelled oligonucleotides to immobilised cells </H3> | ||
| − | <br>E. coli cells expressing each of our 4 zinc finger proteins were immobilised onto glass slides <a href=" | + | <br>E. coli cells expressing each of our 4 zinc finger proteins were immobilised onto glass slides <a href="https://static.igem.org/mediawiki/2015/b/b9/WarwickBacterialProtocolUpdated.pdf">Bacterial Immunofluorescence Protocol</a>. |
<br>Fluorescently labelled oligonucleotides (containing the zinc finger binding domains) were added. | <br>Fluorescently labelled oligonucleotides (containing the zinc finger binding domains) were added. | ||
<br>Binding of the zinc finger proteins to the fluorescent oligonucleotides allows visualisation of the cells by immunofluorescence microscopy. | <br>Binding of the zinc finger proteins to the fluorescent oligonucleotides allows visualisation of the cells by immunofluorescence microscopy. | ||
Revision as of 14:22, 17 September 2015