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− | #season1a, #season2a, #season3a, #season4a, #season5a{ | + | #season1a, #season2a, #season3a, #season4a, #season5a, season6a{ |
width: 90%; | width: 90%; | ||
display:block; | display:block; | ||
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} | } | ||
− | #season1, #season2, #season3, #season4, season5 | + | #season1, #season2, #season3, #season4, season5, season6 |
{ | { | ||
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} | } | ||
− | .close-season1, .close-season2, .close-season3, .close-season4 | + | .close-season1, .close-season2, .close-season3, .close-season4, close-season5, close-season6 |
{ | { | ||
position: relative; | position: relative; | ||
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<li>Planned future experiments </li> | <li>Planned future experiments </li> | ||
</ul> | </ul> | ||
+ | |||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <div style="width: 90%; margin: auto;"> | ||
+ | <div class="hovereffect"> | ||
+ | <img class="img-responsive" src="https://static.igem.org/mediawiki/2016/5/52/TeamKentNotebookSeptember.jpeg" alt=""> | ||
+ | <div class="overlay"> | ||
+ | <h2>September</h2> | ||
+ | <a class="modal_link info" id="season6a" href="#season6">Click here</a> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <!--season6--> | ||
+ | <div id="season6"> | ||
+ | <!--THIS IS IMPORTANT! to close the modal, the class name has to match the name given on the ID class="close-animatedModal" --> | ||
+ | <div class="close-season6"> | ||
+ | <br /> | ||
+ | <br /> | ||
+ | <b>X</b> | ||
+ | </div> | ||
+ | |||
+ | <div class="modal-content"> | ||
+ | <!--Your modal content goes here--> | ||
+ | |||
+ | <h1>September</h1> | ||
+ | <hr class="underline"/> | ||
+ | |||
+ | <h2>August 29 th – September 4 th (Week 10)</h2> | ||
+ | |||
+ | <h2>Day 45</h2> | ||
+ | |||
+ | <ul> | ||
+ | <li>The AFM imaging of samples commenced.</li> | ||
+ | <img style="width: 20%; float: right;" src="https://static.igem.org/mediawiki/2016/0/00/TeamKentSeptemberImage1.jpeg"> | ||
+ | <li>Double digest of PSB1C3 RFP with Sup35nm using Ecor1 and Pst1</li> | ||
+ | <li>PCR using usual protocol (with double digested template DNA) and original long primer on pET23B SecS genes.</li> | ||
+ | <li>Prepared 500ml LB in 2L Baffled flasks for protein expression</li> | ||
+ | </ul> | ||
+ | |||
+ | <h2>Day 46</h2> | ||
+ | |||
+ | <ul> | ||
+ | <li>Repeated agarose gels</li> | ||
+ | <li>New forward primers arrived therefore PCR was done using normal protocol (with annealing temp at 52 degree and double digested template DNA).</li> | ||
+ | <li>We set up 4ml overnight liquid cultures containing double antibiotics, 2 for each Pet23b Sec.S MamT, MamP, and MamX with pEC86 using the same colony.</li> | ||
+ | <li>Made fresh antibiotic samples.</li> | ||
+ | </ul> | ||
+ | |||
+ | <h2>Day 47</h2> | ||
+ | |||
+ | <ul> | ||
+ | <li>Made 1ml glycerol stocks using 500ul of liquid culture for Sec.S MamP, MamT and MamX with Pec86 in BL21’s.</li> | ||
+ | <li>Using the remaining 3.5ml to inoculate 500ml LB in the Baffled flasks.</li> | ||
+ | <li>We grew up the cells and induced with IPTG for protein expression.</li> | ||
+ | <li>Ligation and transformations carried out.</li> | ||
+ | </ul> | ||
+ | |||
+ | <h2>Day 48</h2> | ||
+ | |||
+ | <ul> | ||
+ | <li>We finalised our plans for collaboration with UEA over skype conference. It was agreed that we would carry out some AFM images of their S. oneidensis cells on a variety of different surfaces.</li> | ||
+ | <li>SDS gels carried for MamP SecS, MamX SecS, MamT SecS and stained overnight.</li> | ||
+ | </ul> | ||
+ | |||
+ | <h2>Day 49</h2> | ||
+ | |||
+ | <ul> | ||
+ | <li>De-strained SDS page gels and imaged them.</li> | ||
+ | <li>In preparation, we shipped off AFM sample discs to UEA in Norwich in preparation for collaboration the following week.</li> | ||
+ | </ul> | ||
+ | |||
+ | <h2>September 5 th - September 11 th (Week 11)</h2> | ||
+ | <img style="width: 20%; float: right;" src="https://static.igem.org/mediawiki/2016/0/00/TeamKentSeptemberImage2.jpeg"> | ||
+ | |||
+ | <h2>Day 50</h2> | ||
+ | |||
+ | <ul> | ||
+ | <li>We prepared liquid cultures with all Mam pET23B genes.</li> | ||
+ | <li>Using plasmid maps we designed new primers.</li> | ||
+ | </ul> | ||
+ | |||
+ | <h2>Day 55</h2> | ||
+ | |||
+ | <ul> | ||
+ | <li>Mini-prepped liquid cultures and ran undigested verification gels</li> | ||
+ | <li>Re-attempted PCR</li> | ||
+ | <li>Web design progress for our wiki.</li> | ||
+ | <li>We used the absorbance reading at 280nm and extinction coefficient 8.73 mM -1 cm -1 to estimate the concentration of mam P as 27.4 µM.</li> | ||
+ | <li>We used the absorbance reading at 280 and extinction coefficient 27.32 mM -1 cm -1 to estimate the concentration of mam P as 39.2 µM</li> | ||
+ | </ul> | ||
+ | |||
+ | <h2>Day 56</h2> | ||
+ | |||
+ | <ul> | ||
+ | <li>Using SnapGene plasmid map sequences we designed new SecS primers</li> | ||
+ | <li>We carried out spin columns for protein purification of mamP/T/X</li> | ||
+ | <li>Using the absorbance reading at 280 nm and extinction coefficients, we estimated the concentrations for mamP/X/T for subsequent anaerobic reaction vessels.</li> | ||
+ | <img style="width: 20%; float: right;" src="https://static.igem.org/mediawiki/2016/0/00/TeamKentSeptemberImage3.jpeg"> | ||
+ | <li>Carried out the reaction using purified proteins with iron solutions within anaerobic reaction vessel.</li> | ||
+ | </ul> | ||
+ | |||
+ | <h2>Day 57</h2> | ||
+ | |||
+ | <ul> | ||
+ | <li>Reactions between purified proteins and iron solutions were placed onto EM grids and prepared for imaging (in vivo).</li> | ||
+ | </ul> | ||
+ | |||
+ | <h2>September 12 th – September 18 th (Week 12)</h2> | ||
+ | |||
+ | <ul> | ||
+ | <li>The team logo and banner were designed for review by the team.</li> | ||
+ | <li>We collaborated UEA to AFM image with samples.</li> | ||
+ | <li>We continued making our final constructs.</li> | ||
+ | </ul> | ||
+ | |||
+ | <h2>September 19 th - September 25 th (Week 13)</h2> | ||
+ | |||
+ | <ul> | ||
+ | <li>The team logo and banner were finalised.</li> | ||
+ | <li>We continued writing</li> | ||
+ | <li>AFM imaging of our samples continued.</li> | ||
+ | </ul> | ||
+ | |||
</div> | </div> | ||
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} | } | ||
}); | }); | ||
+ | |||
+ | $("#season6a").animatedModal({ | ||
+ | modalTarget:'season6', | ||
+ | animatedIn:'zoomIn', | ||
+ | animatedOut:'zoomOut', | ||
+ | color:'#330033', | ||
+ | // Callbacks | ||
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+ | console.log("The animation was called"); | ||
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+ | afterOpen: function() { | ||
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Revision as of 23:28, 19 October 2016