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ULB/5 October 2010
From 2010.igem.org
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{{ULB_Header_2}} | {{ULB_Header_2}} | ||
| - | + | '''Biobricks construction''' | |
*PCR to obtain linearised pSB1C3, using linearised pSB1C3 as template, with the protocol from iGEM parts registry. Didn't work. | *PCR to obtain linearised pSB1C3, using linearised pSB1C3 as template, with the protocol from iGEM parts registry. Didn't work. | ||
*PCR on ParD and ParE to have those genes outside of a plasmid (as PCR products). Worked. We used a plasmid containing both genes as template. | *PCR on ParD and ParE to have those genes outside of a plasmid (as PCR products). Worked. We used a plasmid containing both genes as template. | ||
*PCR on FLP, using the sequence we got synthesized as backbone, to add RCF 10 prefix and suffix to the gene. Didn't work. | *PCR on FLP, using the sequence we got synthesized as backbone, to add RCF 10 prefix and suffix to the gene. Didn't work. | ||
| + | *Digestion of Cm and Kan resistance cassettes with RCF10 enzymes in order to insert them in pSB1C3. Worked. | ||
| - | + | '''Module hydrogen''' | |
* We obtained bacteria strains with two deleted genes: one strain that lacked LdhA and FocA and the other one was missing LdhA and PPC. | * We obtained bacteria strains with two deleted genes: one strain that lacked LdhA and FocA and the other one was missing LdhA and PPC. | ||
Latest revision as of 00:08, 28 October 2010
Biobricks construction
- PCR to obtain linearised pSB1C3, using linearised pSB1C3 as template, with the protocol from iGEM parts registry. Didn't work.
- PCR on ParD and ParE to have those genes outside of a plasmid (as PCR products). Worked. We used a plasmid containing both genes as template.
- PCR on FLP, using the sequence we got synthesized as backbone, to add RCF 10 prefix and suffix to the gene. Didn't work.
- Digestion of Cm and Kan resistance cassettes with RCF10 enzymes in order to insert them in pSB1C3. Worked.
Module hydrogen
- We obtained bacteria strains with two deleted genes: one strain that lacked LdhA and FocA and the other one was missing LdhA and PPC.
