Team:Trieste/notebook6

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Week 6

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Suicide System

Holin is defined as a Bglbrick part, that means it doesnít have the standard restriction sites. We transformed the standard Biobrick we needed to use, in Bglbrick by PCR.

The promoter T5CumateOperator was cut EcoRI/SpeI, the fragment was eluted from the gel and then cloned upstream the RBS_B0034, into the pSB1A2 linearized backbone.

We screened the colonies by PCR with the standard VF2 and VR primers and the positive ones were inoculated; we amplified the miniprep by PCR with particular primers with the EcoRI, BglII, BamHI and XhoI restriction sites.

Antibody

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Chassis

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