Difference between revisions of "Team:TNCR Korea/Contribution"

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<h1>Contribution</h1>
 
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<h3>Bronze Medal Criterion #4</h3>
 
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<p><b>Standard Tracks:</b> Participate in the Interlab Measurement Study (to be documented on your InterLab page) and/or improve the characterization of an existing BioBrick Part or Device and enter this information on that part's Main Page in the Registry. The part that you are characterizing must NOT be from a 2017 part number range. Teams who are working on improving the characterization of an existing part should document their experimental design here, along with an explanation for why they chose that part to improve. Data can also be shown here, but it MUST also be documented on the part's Main Page in the Registry.
 
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<b>Special Tracks:</b> Document at least one new substantial contribution to the iGEM community that showcases a project related to BioBricks. This contribution should be central to your project and equivalent in difficulty to making and submitting a BioBrick part.
 
  
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During the use biobrick parts or biological devices (like a promoter), it is very limited to use the EcoR1, SpeI, XbaI, PstI enzyme sites for the insert gene. To compensate for this restriction, we’ve decided to use the primer design used in infusion cloning.
 
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<p> 1. It is difficult to identify whether the Anderson promoter is ligated to the vector since the Anderson promoter is only 33bp. Thus, according to the other teams as well, the Anderson promoter wasn’t ligated into the 3a assembly and amplified. Instead, they’ve redesigned the oligo of the Anderson promoter(5' EcoR1 overhang---- 3' spe1 3'over hang) and used the annealing method. However, TNCR Korea used a different method in hopes of using the anderson promoters that enable gene expression control. If we use the infusion primer design and attach the Anderson promoter at the primer and go through PCR the Anderson promoter will naturally amplify. We can identify this with the PCR product bend test. This is because if the primer doesn’t work, we won’t be able to identify the bend.</p>
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2. Like DPP4, if there are EcoR1, Spe1, Xba1, Pst1 site which are used in 3A assembly, it is read as illegal insert gene which is unable to use. Thus, a step that changes a sequence like point mutation is required which is infusion cloning. Ligation utilized in infusion cloning only requires cutting a vector, not enzyme cut of the insert gene. This is because, insert gene naturally inserts itself into vector while it is amplified by PCR with primer sequence.
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Revision as of 03:47, 2 November 2017

TNCR_KOREA — Digestable Gluten

Parts

Contribution

During the use biobrick parts or biological devices (like a promoter), it is very limited to use the EcoR1, SpeI, XbaI, PstI enzyme sites for the insert gene. To compensate for this restriction, we’ve decided to use the primer design used in infusion cloning.

1. It is difficult to identify whether the Anderson promoter is ligated to the vector since the Anderson promoter is only 33bp. Thus, according to the other teams as well, the Anderson promoter wasn’t ligated into the 3a assembly and amplified. Instead, they’ve redesigned the oligo of the Anderson promoter(5' EcoR1 overhang---- 3' spe1 3'over hang) and used the annealing method. However, TNCR Korea used a different method in hopes of using the anderson promoters that enable gene expression control. If we use the infusion primer design and attach the Anderson promoter at the primer and go through PCR the Anderson promoter will naturally amplify. We can identify this with the PCR product bend test. This is because if the primer doesn’t work, we won’t be able to identify the bend.

2. Like DPP4, if there are EcoR1, Spe1, Xba1, Pst1 site which are used in 3A assembly, it is read as illegal insert gene which is unable to use. Thus, a step that changes a sequence like point mutation is required which is infusion cloning. Ligation utilized in infusion cloning only requires cutting a vector, not enzyme cut of the insert gene. This is because, insert gene naturally inserts itself into vector while it is amplified by PCR with primer sequence.