Difference between revisions of "Team:Hong Kong HKU/projectoverview"
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<h2>2. Synthesis of DNA Nanostructure</h2> | <h2>2. Synthesis of DNA Nanostructure</h2> | ||
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<p>Our two-dimensional DNA nanostructure was synthesized through thermal annealing of the six individual oligonucleotide strands. The hydrogen bonds of the DNA melt at a high temperature in the thermal cycler and upon cooling, the most stable structure, which is our desired nanostructure, is formed. | <p>Our two-dimensional DNA nanostructure was synthesized through thermal annealing of the six individual oligonucleotide strands. The hydrogen bonds of the DNA melt at a high temperature in the thermal cycler and upon cooling, the most stable structure, which is our desired nanostructure, is formed. | ||
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<p>Polyacrylamide gel electrophoresis (PAGE) was carried out to assess the complementary binding of the structure oligonucleotides and the successful formation of the structure. Interaction and binding of oligonucleotides cause gel bands to move up due to higher molecular size. </p> | <p>Polyacrylamide gel electrophoresis (PAGE) was carried out to assess the complementary binding of the structure oligonucleotides and the successful formation of the structure. Interaction and binding of oligonucleotides cause gel bands to move up due to higher molecular size. </p> | ||
| − | <img src="" alt=""> | + | <img src="https://static.igem.org/mediawiki/2017/0/03/HKUPicture3.png" alt=""> |
<h2> 4. Detection of Target </h2> | <h2> 4. Detection of Target </h2> | ||
<p>Fluorometric assay was carried out to evaluate the effectiveness of the DNA nanostructure in detecting the Huntington’s disease miRNA biomarker, Hsa-miR-34b. The two-dimensional DNA nanostructure changes to a three-dimensional structure on detection of target. | <p>Fluorometric assay was carried out to evaluate the effectiveness of the DNA nanostructure in detecting the Huntington’s disease miRNA biomarker, Hsa-miR-34b. The two-dimensional DNA nanostructure changes to a three-dimensional structure on detection of target. | ||
</p> | </p> | ||
| − | <img src="" alt=""> | + | <img src="https://static.igem.org/mediawiki/2017/e/e5/HKUPicture4.png" alt=""> |
<h2>5. Evaluation and Improvement </h2> | <h2>5. Evaluation and Improvement </h2> | ||
<p>Input was received from a number of potential end-users of a diagnostic test made using our DNA nanostructure, for instance through questionnaires sent to medical professionals. All feedback was taken into account and incorporated into the redesigning and improvement of our structure. | <p>Input was received from a number of potential end-users of a diagnostic test made using our DNA nanostructure, for instance through questionnaires sent to medical professionals. All feedback was taken into account and incorporated into the redesigning and improvement of our structure. | ||
</p> | </p> | ||
| − | <img src="" alt=""> | + | <img src="https://static.igem.org/mediawiki/2017/2/22/HKUPicture5.png" alt=""> |
<h2>6. Synthesis of Biobricks</h2> | <h2>6. Synthesis of Biobricks</h2> | ||
<p>Finally, five biobricks were produced, which can produce the five single-stranded DNA oligonucleotides of our structure respectively. The ssDNA can come together to form our functional tetrahedral structure in the presence of the specific target. | <p>Finally, five biobricks were produced, which can produce the five single-stranded DNA oligonucleotides of our structure respectively. The ssDNA can come together to form our functional tetrahedral structure in the presence of the specific target. | ||
</p> | </p> | ||
| − | <img src="" alt=""> | + | <img src="https://static.igem.org/mediawiki/2017/6/67/HKUPicture6.png" alt=""> |
Revision as of 08:21, 29 October 2017
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Project Overview
1. Planning and design
After extensive brainstorming and research, our DNA nanostructure was designed with the help of the software, Tiamat.
2. Synthesis of DNA Nanostructure
Our two-dimensional DNA nanostructure was synthesized through thermal annealing of the six individual oligonucleotide strands. The hydrogen bonds of the DNA melt at a high temperature in the thermal cycler and upon cooling, the most stable structure, which is our desired nanostructure, is formed.
3. Observation of DNA Structure Formation
Polyacrylamide gel electrophoresis (PAGE) was carried out to assess the complementary binding of the structure oligonucleotides and the successful formation of the structure. Interaction and binding of oligonucleotides cause gel bands to move up due to higher molecular size.
4. Detection of Target
Fluorometric assay was carried out to evaluate the effectiveness of the DNA nanostructure in detecting the Huntington’s disease miRNA biomarker, Hsa-miR-34b. The two-dimensional DNA nanostructure changes to a three-dimensional structure on detection of target.
5. Evaluation and Improvement
Input was received from a number of potential end-users of a diagnostic test made using our DNA nanostructure, for instance through questionnaires sent to medical professionals. All feedback was taken into account and incorporated into the redesigning and improvement of our structure.
6. Synthesis of Biobricks
Finally, five biobricks were produced, which can produce the five single-stranded DNA oligonucleotides of our structure respectively. The ssDNA can come together to form our functional tetrahedral structure in the presence of the specific target.
