Part:BBa_K4268000:Experience

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Applications of BBa_K4268000

The part was received, resuspended, and cloned into the level 0 Golden Gate vector, PSB1C00. The ligation was then transformed into DH5&alpha cells. After overnight growth, white colonies (indicative of a replacement of the RFP reporter cassette) were subjected to screening for the correct insert using colony PCR with the VF/VR primer pair.

Figure 1: Colony PCR of five colonies (clones) suspected of coating the insert Head-tail connector insert. The insert length is 1573bp and the predicted size of the PCR product using VF/VR primers is 1739 bp.

The gel indicates that all five colonies are likely to contain the correct insert and thus, the Head-Tail Connector was successfully cloned into a Level O Golden Gate Assembly basic part.

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