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− | <h1>Parts</h1> | + | <h1>Parts</h1> |
+ | <p>This year we produced 5 biobricks each of which produce ssDNA which can come together to form a functional tetrahedron structure in presence of its target. The inserts were ordered as gBlocks fragments from Integrated DNA technologies As shown in the diagram, the insert can be divided into 5 parts:</p> | ||
+ | <ol> | ||
+ | <li>Biobrick prefix and suffix </li> | ||
+ | <li>ProD - This is the Promoter for our biobrick. The promoter allows for high expression of our downstream product </li> | ||
+ | <li>ssDNA sequence: This sequence is complementary to the ncRNA sequence that will be produced.</li> | ||
+ | <li>HTBS: HIV reverse transcriptase binding site </li> | ||
+ | <li>BBa_B0054 : Strong Terminator </li> | ||
+ | </ol> | ||
− | <p> | + | <p>We expect that upon transcription a ncRNA will be produced with an HTBS sequence that will allow HIV reverse transcriptase to bind the ncRNA and produce the desired DNA sequence. Upon addition of RNase H and RNase A the DNA: RNA duplex and the degradation can occur respectively. </p> |
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− | + | <p> As shown before the Pre-tetra and the Tetra is generated using five nucleotide sequence and we have made a total of 5 biobricks, one biobrick for each. </p> | |
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− | + | <center><img src="https://static.igem.org/mediawiki/parts/b/ba/Plasmidconstruct1.png" alt="plasmidconstruct" style="width:50%; height:50%" > | |
− | + | <h4>Figure 1: Structure of insert</h4> | |
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− | + | <img src="https://static.igem.org/mediawiki/2017/7/74/Target_Vector_History.png" alt="insertadd" style="width:50%; height:50%"> | |
− | + | <h4>Figure 2: Production of biobrick</h4> | |
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− | + | <img src="https://static.igem.org/mediawiki/2017/0/0c/Webp.net-resizeimageHKU.png" alt="New gel" style="width:50%; height:50%"> | |
− | + | <h4>Figure 3: Polyacrylamide gel electrophoresis (PAGE) of individual oligonucleotides of our DNA nanostructure</h4> | |
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Latest revision as of 16:34, 1 November 2017
Parts
This year we produced 5 biobricks each of which produce ssDNA which can come together to form a functional tetrahedron structure in presence of its target. The inserts were ordered as gBlocks fragments from Integrated DNA technologies As shown in the diagram, the insert can be divided into 5 parts:
- Biobrick prefix and suffix
- ProD - This is the Promoter for our biobrick. The promoter allows for high expression of our downstream product
- ssDNA sequence: This sequence is complementary to the ncRNA sequence that will be produced.
- HTBS: HIV reverse transcriptase binding site
- BBa_B0054 : Strong Terminator
We expect that upon transcription a ncRNA will be produced with an HTBS sequence that will allow HIV reverse transcriptase to bind the ncRNA and produce the desired DNA sequence. Upon addition of RNase H and RNase A the DNA: RNA duplex and the degradation can occur respectively.
As shown before the Pre-tetra and the Tetra is generated using five nucleotide sequence and we have made a total of 5 biobricks, one biobrick for each.
Figure 1: Structure of insert
Figure 2: Production of biobrick
Figure 3: Polyacrylamide gel electrophoresis (PAGE) of individual oligonucleotides of our DNA nanostructure