-Speeding Up-

Extracted plasmid from E.coli transformated BBa_J04450,

Refined by phenol chloroform mixture and ethanol precipitation,

Added restriction enzyme,

HNS(wt)→X,S

BBa_J04450→X,S

-Targetting-

@plasmid extraction and DNA refine J23100-P0440

@colony PCR J23100-K317037 and J23100-P0440

@electrophoresis
J23100-K317037: find binds in 29 and 31
J23100-P0440: find binds almost all

@preculture
J23100-K317037 and J23100-P0440 which find binds.

-Visualization-

-Ligation

Promoter - suppressed expression system