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Team:Edinburgh/Experiments
From 2011.igem.org
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For our tests, we attempted to make a construct containing a Plac-LacZ part, as well as EYFP. BioSandwich inserts short spacers between each part to be assembled. Note that after spacer2 we expect to see some extra bases before we reach the RFC10 suffix. We can call these extra bases the "stuffing". So, the expected sequence is: | For our tests, we attempted to make a construct containing a Plac-LacZ part, as well as EYFP. BioSandwich inserts short spacers between each part to be assembled. Note that after spacer2 we expect to see some extra bases before we reach the RFC10 suffix. We can call these extra bases the "stuffing". So, the expected sequence is: | ||
| - | : Vector -- spacerT7 -- Plac,LacZ -- spacer1 -- EYFP -- spacer2 -- stuffing | + | : '''Vector -- spacerT7 -- Plac,LacZ -- spacer1 -- EYFP -- spacer2 -- stuffing''' |
Successful assemblies are expected to generate blue colonies that fluoresce yellow under a blue light. We tried several assembly methods, all of which generated many colonies with the correct phenotype, but also many colonies with an incorrect phenotype... | Successful assemblies are expected to generate blue colonies that fluoresce yellow under a blue light. We tried several assembly methods, all of which generated many colonies with the correct phenotype, but also many colonies with an incorrect phenotype... | ||
Revision as of 11:15, 18 August 2011
BioSandwich
The BioSandwich system is outlined on its own page. BioSandwich creates parts with homologous ends, but there are a number of ways in which the final assembly could be carried out.
For our tests, we attempted to make a construct containing a Plac-LacZ part, as well as EYFP. BioSandwich inserts short spacers between each part to be assembled. Note that after spacer2 we expect to see some extra bases before we reach the RFC10 suffix. We can call these extra bases the "stuffing". So, the expected sequence is:
- Vector -- spacerT7 -- Plac,LacZ -- spacer1 -- EYFP -- spacer2 -- stuffing
Successful assemblies are expected to generate blue colonies that fluoresce yellow under a blue light. We tried several assembly methods, all of which generated many colonies with the correct phenotype, but also many colonies with an incorrect phenotype...
