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From 2011.igem.org

We ordered our genes. We optimized codon usage because of gene taken from E. cloacae.

Codon Usage Then we did restriction digestion and gel electrophoresis.
We repeat this step a lot because of failuire.I put right result

We used standard gel purification kit Our ligation style is really similar to iGEM's one.
Our one;

insert volume(µl):X

Cut Vector Volume(µl):X

DEPC-water(µl):8,5-2X

T4 DNA ligase(µl):0,5

Ligation Buffer(µl):1

Total Volume(µl):10

We transform our plasmid into E.coli with a protocol