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<h2>Week 1: Ideation</h2> | <h2>Week 1: Ideation</h2> | ||
− | <p style="font-family: 'Lato'; font-size:15px"> | + | <p style="font-family: 'Lato'; font-size:15px">During the first week, we tried to come up with ideas that we can work on. After long discussions and much debate, we finally decided to work on an issue that |
− | + | we think is quite relevant not only in our country but also in many developing and | |
− | we think is quite relevant not only in our country but also in | + | developed countries — that of stinking public toilets. For our |
− | developed countries | + | inspiration and project summary, see the <a href="https://2017.igem.org/Team:ICT-Mumbai/Description"><mark>project description</mark></a>.</p> |
− | + | ||
− | inspiration and project summary see <a href="https://2017.igem.org/Team:ICT-Mumbai/Description">project description | + | |
</div> | </div> | ||
</div> | </div> | ||
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<h2>Week 2: Ground Reality</h2> | <h2>Week 2: Ground Reality</h2> | ||
− | <p style="font-family: 'Lato'; font-size:15px">During this week, we decided to approach different | + | <p style="font-family: 'Lato'; font-size:15px">During this week, we decided to approach different NGOs and organizations that are |
involved in promoting and providing better sanitation. Sulabh International is an India-based | involved in promoting and providing better sanitation. Sulabh International is an India-based | ||
NGO that promotes and provides better sanitation by building public pay and use toilets and | NGO that promotes and provides better sanitation by building public pay and use toilets and | ||
− | community toilets | + | community toilets. To understand and evaluate the feasibility |
− | of our idea and to understand the ground reality we decided to meet with | + | of our idea and to understand the ground reality, we decided to meet with a representative of Sulabh |
− | International. | + | International. Details of our interaction with this NGO can be found on our <a href="https://2017.igem.org/Team:ICT-Mumbai/HP/Gold_Integrated"><mark>human practices</mark></a> page.</p> |
<p></p> | <p></p> | ||
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<p style="font-family: 'Lato'; font-size:15px">Week 3, and we step into the lab!<br><br> | <p style="font-family: 'Lato'; font-size:15px">Week 3, and we step into the lab!<br><br> | ||
− | During the first week in the lab, we learnt about the techniques and the principles behind | + | During the first week in the lab, we learnt about the techniques and the principles behind them. It marked our first foray into the world of synthetic biology and it was great fun learning and performing these experiments.<br> |
− | 1.Wet lab begins! We learn about different kinds of growth media, composition and their significance.<br> | + | 1. Wet lab begins! We learn about different kinds of growth media, composition and their significance.<br> |
− | 2.Introduction to agarose gel electrophoresis and DNA quantification methods.<br> | + | 2. Introduction to agarose gel electrophoresis and DNA quantification methods.<br> |
− | Additionally, we learnt the precautions to follow while handling ethidium bromide containing gels and | + | Additionally, we learnt the precautions to follow while handling ethidium bromide containing gels and their disposal procedures. |
− | 3.We also kicked off our very first step towards establishing our proof of concept | + | 3. We also kicked off our very first step towards establishing our proof of concept — extracting genomic DNA from <i>Escherichia coli</i> BW25113. We checked the size of the extracted product by agarose gel electrophoresis.<br><br> |
<img src="https://static.igem.org/mediawiki/2017/4/4b/ICT-Mumbai_notebook_gdna_image.png" alt=""><br> | <img src="https://static.igem.org/mediawiki/2017/4/4b/ICT-Mumbai_notebook_gdna_image.png" alt=""><br> | ||
− | 5ul and 10ul of the extracted products were loaded in lanes 1,3 and 2,4 respectively. | + | 5ul and 10ul of the extracted products were loaded in lanes 1, 3 and 2, 4 respectively. |
</p> | </p> | ||
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<h2>Week 4:The idea!</h2> | <h2>Week 4:The idea!</h2> | ||
− | <p style="font-family: 'Lato'; font-size:15px">Glutamine synthetase (<i>glnA</i>), smaller and bigger subunits of glutamate synthetase (<i>gltD</i> and <i>gltB</i> respectively) were our genes of primary interest. All the three genes of interest were amplified from <i> E. coli </i> | + | <p style="font-family: 'Lato'; font-size:15px">Glutamine synthetase (<i>glnA</i>), and the smaller and bigger subunits of glutamate synthetase (<i>gltD</i> and <i>gltB</i>, respectively) were our genes of primary interest. All the three genes of interest were amplified from <i>E. coli</i> BW25113 genomic DNA using polymerase chain reaction (PCR). PCR products were loaded and run on 1% agarose gel to check for the correct sizes of these three genes (<i>glnA</i>: 1640 bp, <i>gltB</i>: 4462 bp, <i>gltD</i>: 1420 bp). Among the three, we were successful in obtaining two genes — <i>glnA</i> and <i>gltD</i>. We repeated PCR for <i>gltB</i> for the second time by adding DMSO. This time too we could not see bands of correct size. We again carried out PCR using different annealing temperatures and the product obtained with 62°C as annealing temperature gave good bands. This was further gel purified and used as a template to carry out another PCR for the same gene. We could not see bands of correct size again.</p> |
− | + | ||
− | + | ||
</div> | </div> | ||
</div> | </div> |
Revision as of 22:09, 1 November 2017