Team:Paris Bettencourt/Notebook/Trojan Horse/Wednesday 21st August.html

Trojan Horse

Place your twit here

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