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| | ===Applications of BBa_K540000=== | | ===Applications of BBa_K540000=== |
| − | <html>
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| − | <div style="text-align:center; float:right; margin-left:30px; margin-top:-20px;">
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| − | <img src="http://i.imgur.com/g7iTW.png" style="width:340px; height:255px; margin-top: 70px; margin-bottom: 5px; position: relative;"/>
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| − | </div>
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| − | <p>
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| − | A preliminary search on the best condition for use of this part was carried out at first.
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| − | </p><br>
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| − | <p>
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| − | <FONT COLOR="green"><b>Antibiotic response</b></FONT><br>
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| − | Two 24 wells plates have been seeded with the same strains, in LB/2 medium. Ampicilline for S18 and Cm have been added only in the second plate.
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| − | The results have examined after 24 hours of incubation 30°C.
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| − | </p><br>
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| − | <p><i>Qualitative approach</i><br>
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| − | As can be seen on the right, adding ampicilline boosts the adherence of the strain, and makes it more cobalt responsive.
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| − | </p>
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| − | <div style="text-align:center; float:left; margin-left:30px; margin-top:-20px;">
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| − | <img src="http://i.imgur.com/wnOY5.png" style="width:525px; height:350px; margin-top: 100px; margin-bottom: 5px; position: relative;"/>
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| − | </div>
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| − |
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| − | <p>
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| − | <br/><br/>
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| − | <i>Quantitative approach</i><br>
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| − | Furthermore, the adherence level was determined quantitatively using OD measurements. Resulting graphs are shown on the left.</p><br>
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| − | <p>
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| − | P1 indicates the first plate, without antibiotics, while P2 indicates the second plate, with antibiotics.</p><br>
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| − | <p>
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| − | <b>Conclusion</b>: it can clearly be seen that while the growth of the bacteria are similar, as they have similar total OD, adding the antibiotics makes them a lot more adherent, and enable the cobalt-response.</p><br>
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| − | <p>
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| − | This can be explained by the fact that without antibiotics, the bacteria will tend to lose the part, and thus become unresponsive to cobalt.
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| − | </p><br>
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| − | <div style="text-align:center; float:right; margin-left:30px; margin-top:-20px;">
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| − | <img src="http://i.imgur.com/zIwBU.png" style="width:402px; height:350px; margin-top: 60px; margin-bottom: 5px; position: relative;"/>
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| − | </div>
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| − | <br><br><br><br>
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| − | <p>
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| − | <FONT COLOR="green"><b>Medium influence</b></FONT><br>
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| − | During adherence tests, bacteria can be grown in LB/2 medium (50% Lb, 50% sterile water) or in M63G medium (100mL M63, 1ml LB, 1 ml Glucose 20%).
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| − | </p><br>
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| − | <p>
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| − | In order to choose the better medium, two strains have been seeded in the 2 different media, with a concentration of 10^7 cells/mL. The first is an adherent strain PHL818, the second is a non-adherent strain NM522.
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| − | Plates have been incubated at 30°C during 48 hours.
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| − | </p><br>
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| − | <p>
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| − | Results which are show on the right are a compilation of repetitions of this experiment.
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| − | </p><br>
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| − | <p>
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| − | <b>Conclusion</b>: a greater biofilm formation is observed using the M63G medium.
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| − | <div style="text-align:center; float:left; margin-right:20px; margin-top:-20px;">
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| − | <img src="http://i.imgur.com/p0pEe.png" style="width:308px; height:447px; margin-top: 110px; margin-bottom: 5px; position: relative;"/>
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| − | </div>
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| − | <br><br><br><br><br>
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| − | <p>
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| − | <FONT COLOR="green"><b>EDTA cleaning influence</b></FONT><br>
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| − | Plates were washed with 4mM EDTA, and rinsed with water before growing bacteria. This aims to reduce the influence of any cobalt or nickel pre-existing in the plates, that might change the actual cobalt concentration in the plates.
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| − | </p><br>
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| − |
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| − | <p>
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| − | Results are on the left. As can be seen, treating with EDTA beforehand increases the adherence of the strain.
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| − | </p><br>
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| − | <p>
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| − | The stable OD proves that differences observed are not due to a difference in growth.
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| − | </p><br>
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| − | <p>
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| − | <b>Conclusion</b>: When using this part, it is preferable to wash the plates beforehand with EDTA (or to use metal-free plastic)
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| − | </p>
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| − | <br><br><br><br><br><br><br><br><br><br><br><br>
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| − | <p>
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| − | <FONT COLOR="red"><b>Conclusion about the preliminary tests of the part</b></FONT><br>
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| − | From these experiments, we can conclude that the best conditions to grow bacteria transformed with this part are :
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| − | <ul>
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| − | <li>Using a selective antibiotic</li>
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| − | <li>M63G medium (100 mL M63 + 1 mL LB + 1 mL Glucose 20%).</li>
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| − | <li>EDTA washing beforehand</li>
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| − | <li>48h growth time</li>
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| − | </ul>
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| − | </p><br>
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| − | <p>
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| − | These conditions are the one used in the characterization described on the main page.
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| − | </p><br>
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| − | <br/><br/><br/><br/><br><br><br><br>
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| | | | |
| | ===User Reviews=== | | ===User Reviews=== |
This experience page is provided so that any user may enter their experience using this part.
Please enter
how you used this part and how it worked out.
UNIQ666d9d40587450c7-partinfo-00000000-QINU
UNIQ666d9d40587450c7-partinfo-00000001-QINU
