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Team:Groningen/Template/MODULE/Notebook/characterisation/week6
From 2014.igem.org
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| - | Testing the constructs RBS-NisA-double terminator (BBa_K1365556) and RBS-Superfolded GFP-double terminator (BBa_K1365555) with some different promoter: P2, PLas, and two constitutive promoters ( | + | Testing the constructs RBS-NisA-double terminator (<a href="http://parts.igem.org/Part:BBa_K1365556">BBa_K1365556</a>) and RBS-Superfolded GFP-double terminator (<a href="http://parts.igem.org/Part:BBa_K1365555">BBa_K1365555</a>) with some different promoter: P2, PLas, and two constitutive promoters (<a href="http://parts.igem.org/Part:BBa_J23101">BBa_J23101</a> and <a href="http://parts.igem.org/Part:BBa_J23115">BBa_J23115</a>), and inserting these constructs in pIL253A (<a href="http://parts.igem.org/Part:BBa_K1365301">BBa_K1365301</a>) vector |
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Revision as of 18:02, 17 October 2014
October 6 - October 12
Testing the constructs RBS-NisA-double terminator (BBa_K1365556) and RBS-Superfolded GFP-double terminator (BBa_K1365555) with some different promoter: P2, PLas, and two constitutive promoters (BBa_J23101 and BBa_J23115), and inserting these constructs in pIL253A (BBa_K1365301) vector
The ligations of the different constructs were performed overnight, after which the ligated plasmids were transformed to L. lactis NZ9000, and finally a colony PCR was performed with several colonies of each construct
The construct with the P2 and PLas did not show any bands on the gel, therefore further steps were only done with the constructs containing the two constitutive promoters
The same constructs with the promoters from the promoter collection of Uppsala (see September 19 till October 5) were subjected to ligation again, but this time the vector pIL235A was used instead of pSB1K3
These constructs were transformed to L. lactis NZ9000, and plated on M17 agar with erythromycin
