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Team:NTNU Trondheim/LacI2
From 2011.igem.org
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When testcutting the constructs containing BBa_K292006 we got some unexpected results. | When testcutting the constructs containing BBa_K292006 we got some unexpected results. | ||
Therefore we decided to digest the brick with three differnt enzymes with restriction sites | Therefore we decided to digest the brick with three differnt enzymes with restriction sites | ||
| - | in the given biobrick-sequence in addition to EcoR1 and PstI | + | in the given biobrick-sequence in addition to EcoR1 and PstI (for looking at the biobrick length) |
| - | + | Except from the sample cut with EcorR1 and PstI, the samples were also cut with BglI wich has a recognition site | |
| - | Except from the sample cut with EcorR1 and PstI, the samples were also cut with BglI | + | |
in the pSB1A2 backbone. | in the pSB1A2 backbone. | ||
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| + | [[File:Test_LacI2_080811_exp_wiki.jpg|thumb|LacI2 plasmid cut with EcoR1 + PstI, BglI + BclI, BglI + EcoRV, BglI + BanII]] | ||
Overwiev of restrctions and expected fragmentlengths: | Overwiev of restrctions and expected fragmentlengths: | ||
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| - | + | As we can see on the resulting gel none of the restriction sites inn the LacI2 fragment appear to be present. In additon the LacI2 insert looks smaller than it's supposed to be. This indicates that the biobrick sequence does not match the | |
| - | + | sequence given in the parts registry. | |
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{{:Team:NTNU_Trondheim/NTNU_footer}} | {{:Team:NTNU_Trondheim/NTNU_footer}} | ||
Latest revision as of 09:34, 17 September 2011
LacI2 (BBa_K292006)
We found and orderd this part from the parts registry. Since it had an RBS and double terminator it would save us from some cloning work.
When testcutting the constructs containing BBa_K292006 we got some unexpected results. Therefore we decided to digest the brick with three differnt enzymes with restriction sites in the given biobrick-sequence in addition to EcoR1 and PstI (for looking at the biobrick length) Except from the sample cut with EcorR1 and PstI, the samples were also cut with BglI wich has a recognition site in the pSB1A2 backbone.
Overwiev of restrctions and expected fragmentlengths:
| Restriction enzymes | Expected lengths (bp) |
|---|---|
| EcoR1 + PstI | 2053 (backbone) and 1303 (biobrick) |
| BglI + BclI | 2032 + 1324 |
| BglI + EcoRV | 1760 + 1596 |
| BglI + BanII | 1835 + 1521 |
As we can see on the resulting gel none of the restriction sites inn the LacI2 fragment appear to be present. In additon the LacI2 insert looks smaller than it's supposed to be. This indicates that the biobrick sequence does not match the sequence given in the parts registry.
