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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 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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| − | <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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| − |
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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:30px; 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><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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| − | <br/><br/><br/><br/><br><br><br><br>
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| | ===User Reviews=== | | ===User Reviews=== |
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| | <!-- Template for a user review | | <!-- Template for a user review |
| | {|width='80%' style='border:1px solid gray' | | {|width='80%' style='border:1px solid gray' |
| − | |- | + | |-Team |
| | |width='10%'| | | |width='10%'| |
| | <partinfo>BBa_K540000 AddReview number</partinfo> | | <partinfo>BBa_K540000 AddReview number</partinfo> |
| − | <I>Username</I> | + | <I>User</I> |
| | |width='60%' valign='top'| | | |width='60%' valign='top'| |
| − | Enter the review inofrmation here.
| + | |
| | |}; | | |}; |
| | <!-- End of the user review template --> | | <!-- End of the user review template --> |
| | <!-- DON'T DELETE --><partinfo>BBa_K540000 EndReviews</partinfo> | | <!-- DON'T DELETE --><partinfo>BBa_K540000 EndReviews</partinfo> |
| | + | |
| | + | {|width='80%' style='border:1px solid gray' |
| | + | |- |
| | + | |width='10%'| |
| | + | <partinfo>BBa_I0500 AddReview 5</partinfo> |
| | + | <I>UCL iGEM 2012</I> |
| | + | |width='60%' valign='top'| |
| | + | |
| | + | UCL iGEM 2012 have removed the original cobalt inducible promoter to create a stand alone csgBAEFG operon with rbs [https://parts.igem.org/wiki/index.php?title=Part:BBa_K729003 BBa_K729003]. We have also designed two additional variations on the part, with a [https://parts.igem.org/wiki/index.php?title=Part:BBa_K729017 T7] and [https://parts.igem.org/wiki/index.php?title=Part:BBa_K729018 constitutive promoter]. |
| | + | |
| | + | |} |
| | + | {|width='80%' style='border:1px solid gray' |
| | + | |- |
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.
UNIQ2ebd3c918ac45ed2-partinfo-00000000-QINU
UNIQ2ebd3c918ac45ed2-partinfo-00000001-QINU
