User:Karmella Haynes/Notebook/BioBrick cloning/2013/03/20: Difference between revisions
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'''Assemblies''' | '''Assemblies''' | ||
* New strategy for mammalian vector: | * New strategy for mammalian vector: | ||
** Insert K-XbaI-NLS-6his-stop into the puro vector that has a deleted promoter. This way, the vector can be customized to carry different promoters. | ** Insert K-XbaI-NLS-6his-stop into the puro vector that has a deleted promoter (MV2, puro). This way, the vector can be customized to carry different promoters. | ||
** Insertion of the ds oligo should destroy the vector's XbaI site | ** Insertion of the ds oligo should destroy the vector's XbaI site | ||
** Will need to check inserts for copy number and orientation | ** Will need to check inserts for copy number and orientation | ||
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* '''Oligo annealing''' | * '''Oligo annealing''' | ||
* See [http://openwetware.org/wiki/User:Karmella_Haynes/Notebook/BioBrick_cloning/2013/03/19 03/19/13] for details about the oligos and reaction set-up | |||
* Redo oligo annealing with 99.9 and 97.9°C starting temperatures... | * Redo oligo annealing with 99.9 and 97.9°C starting temperatures... | ||
Revision as of 19:29, 20 March 2013
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03/20/13
Assemblies
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