Difference between revisions of "Part:BBa K4268000:Experience"
 
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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.
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The part was received, resuspended, and cloned into the level 0 Golden Gate vector, PSB1C00. The ligation was then transformed into DH5α 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.
  
  
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[[File: T-suny-oneonta-t7-head-tail connector colony PCR figure.jpg|200px|border|center|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 when using VF/VR primers is 1739 bp.]]
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[[File: T-suny-oneonta-t7-head tail connector colony PCR figure.png|500px|thumb|center|Figure 1: Colony PCR of five colonies (clones) suspected of containing the Head-tail connector insert. The insert length is 1573bp and the predicted size of the PCR product when using VF/VR primers is 1878 bp.]]
  
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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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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 0 Golden Gate Assembly basic part.
  
  
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Latest revision as of 18:21, 8 October 2022


The part was received, resuspended, and cloned into the level 0 Golden Gate vector, PSB1C00. The ligation was then transformed into DH5α 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 containing the Head-tail connector insert. The insert length is 1573bp and the predicted size of the PCR product when using VF/VR primers is 1878 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 0 Golden Gate Assembly basic part.


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