Difference between revisions of "Part:BBa K4268010:Experience"
 
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Level 0 parts BBa_R0085, BBa_B0034, BBa_K4268002 (Capsid Protein), and BBa_B0015 were cloned into the level 1 Golden Gate vector, PSB1K02. 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.
 
Level 0 parts BBa_R0085, BBa_B0034, BBa_K4268002 (Capsid Protein), and BBa_B0015 were cloned into the level 1 Golden Gate vector, PSB1K02. 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-Level 1 capsid protein colony PCR figure.png|500px|thumb|center|Figure 1: Colony PCR of five colonies (clones) suspected of coating the insert Capsid Protein insert. The insert length is 720bp and the predicted size of the PCR product when using VF/VR primers is 1025 bp.]]
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[[File: T-suny-oneonta-t7-Level 1 capsid protein colony PCR figure.png|500px|thumb|center|Figure 1: Colony PCR of five colonies (clones) suspected of containing the Capsid Protein insert. The insert length is 906bp (including the fusion sites) and the predicted size of the PCR product when using VF/VR primers is 1211 bp.]]
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The gel indicates that all five colonies are likely to contain the correct insert and thus, the Level 1 Capsid Protein Transcriptional Unit was successfully cloned.
  
  

Latest revision as of 18:19, 8 October 2022


Level 0 parts BBa_R0085, BBa_B0034, BBa_K4268002 (Capsid Protein), and BBa_B0015 were cloned into the level 1 Golden Gate vector, PSB1K02. 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 Capsid Protein insert. The insert length is 906bp (including the fusion sites) and the predicted size of the PCR product when using VF/VR primers is 1211 bp.

The gel indicates that all five colonies are likely to contain the correct insert and thus, the Level 1 Capsid Protein Transcriptional Unit was successfully cloned.


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