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Team:Copenhagen/Results
From 2012.igem.org
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<table id="graa" cellpadding=20px><td width="890px" height="100%" valign="top" ><p align="justify"><h2>ProC and YFI</h2> | <table id="graa" cellpadding=20px><td width="890px" height="100%" valign="top" ><p align="justify"><h2>ProC and YFI</h2> | ||
| - | We performed USER cloning and managed to assemble ProC and YF1 in the pSB1C3 backbone. This has been submitted as a BioBrick. <br><br> | + | We performed USER cloning and managed to assemble ProC and YF1 in the pSB1C3 backbone. This has been submitted as a BioBrick. <br> For further information please visit the Parts page. <br><br> |
<h2>Lux Cassette</h2> | <h2>Lux Cassette</h2> | ||
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We contacted Thomas Knight from the iGEM headquarter who kindly provided us with a Lux Cassette. This contained one of the “banned” Xbal restriction sites, which we have removed and the Lux cassette is submitted as a BioBrick. | We contacted Thomas Knight from the iGEM headquarter who kindly provided us with a Lux Cassette. This contained one of the “banned” Xbal restriction sites, which we have removed and the Lux cassette is submitted as a BioBrick. | ||
We have transformed the Lux cassette from Thomas Knight into E.cloni® 5α (purchased from Lucigen). | We have transformed the Lux cassette from Thomas Knight into E.cloni® 5α (purchased from Lucigen). | ||
| - | As a control we exploited that the gene cassette had a LacI promotor and we confirmed, after adding IPTG, that the Lux cassette is able to make the bacteria bioluminescent. <br><br> | + | As a control we exploited that the gene cassette had a LacI promotor and we confirmed, after adding IPTG, that the Lux cassette is able to make the bacteria bioluminescent. <br> For further information please visit the Parts page. <br><br> |
<h2>Experiences with USER cloning</h2> | <h2>Experiences with USER cloning</h2> | ||
Revision as of 10:54, 26 September 2012
ProC and YFIWe performed USER cloning and managed to assemble ProC and YF1 in the pSB1C3 backbone. This has been submitted as a BioBrick.For further information please visit the Parts page. Lux CassetteAfter trying to amplify the Lux cassette from the BioBrick plate several times, we checked the sequence (from sequencing) in the parts registry using BLAST and found that it was an E.coli surface protein and not the Lux cassette. We contacted Thomas Knight from the iGEM headquarter who kindly provided us with a Lux Cassette. This contained one of the “banned” Xbal restriction sites, which we have removed and the Lux cassette is submitted as a BioBrick. We have transformed the Lux cassette from Thomas Knight into E.cloni® 5α (purchased from Lucigen). As a control we exploited that the gene cassette had a LacI promotor and we confirmed, after adding IPTG, that the Lux cassette is able to make the bacteria bioluminescent.For further information please visit the Parts page. Experiences with USER cloningWe have found, that in order to make USER cloning work, it is very important that the molecular concentration of the parts, that are to be assembled, is as equal as possible. This means that a small piece must be of lower concentration in order to assemble it to a larger piece so the number of each piece is approximately the same. For ProC (35 bp) and YF1 (1131bp) a x10.000 dilution of ProC was necessary in order to assemble the two pieces. This very problem has made it hard to assemble our construct since we have pieces ranging from 35bp to almost 6000bp. |
