Team:Paris Bettencourt/Notebook/Trojan Horse/Wednesday 21st August.html
From 2013.igem.org
Trojan Horse
ASDF21st August
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Results of assemblies (Colonies on the transformation plates):
sRNA –Kan into Litmus by Gibson: Success
sRNA – Kan into Litmus by biobrick assembly: Success
sRNA – Kan into pUC18 by Gibson: Failure
sRNA – Kan into pUC18 by biobrick assembly: Success
Colony PCR to check if sRNA-KAN is in our Litmus phagemid: positive band between 400 and 500: (expected length 419bp)
Check the design of primers to clone out Kan => problem => redesign them
Design of the primers to sequence all the cloning
Launch 3 colonies of each successful cloning
Glycerol and mini prep of all the launched colonies / need 3 before we have
Construction of target strain
Electroporation of pCOLA in MGZ1 F+
Construction of weapon strains
Electroporation of litmus-GFP-sRNA in XL1 Blue-M13KO7-Helper
Electroporation of Litmus-GFP-sRNA and M13K07 in MGZ1 F+
Chemical transformation of minigene+vector-sRNA anti Kan in NEB turbo