Difference between revisions of "Team:BIT/Interlab Study"

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<li><a href="https://2016.igem.org/Team:BIT/Judging_Form" ><span class="style1">Judging</span></a></li>
 
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Chinese Premier Li Keqiang delivers a keynote speech on October 11 at the opening ceremony of the Fifth Ministerial Conference of The Forum for Economic and Trade Cooperation Between China and Portuguese-speaking Countries in Macao. [Photo/Xinhua]<br><br></p>
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The objective of this year’s iGEM Interlab study is to quantify expression of five different reporter constructs. These constructs have GFP under the control of different promoter and ribosome binding sequences (RBS). Our Interlab work was carried out with the DH5-alpha as an expression host, and the Thermo Scientific Varioskan Flash spectral scanning as the equipment. We transfer the five reporter constructs into E.coli cells separately. And we measure the fluorescence data and optical density data with spectral scanning machine every 1 hour during a 6 hours cultivation. <br><br></p>
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<h2><br><br><br><br>Methods</h2>
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Premier Li Keqiang announced 18 measures on Tuesday in Macao to boost ties with Portuguese-speaking countries in fields such as production capacity, people-to-people exchanges and maritime development.<br><br>
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We use endonuclease (EcoRI and PstI, XbaI and PstI) and T4 DNA ligase (NEB) to construct them. Finally, the ligation product was introduced into the pSB1C3. All constructs used were transformed into DH5α cells.<br><br>
 
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The measures will be implemented in the next three years, and the Chinese government will support the Macao Special Administrative Region in establishing a financial service program, cultural exchange centers, bilingual training bases and juvenile innovation centers for exchanges with these countries, the premier said.<br><br>
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The cultivation of our bacteria was performed in tubes filled with 7 mL LB liquid medium. The cultures were kept at 37 °C and 110 rpm in shaker. <br><br>
 
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China resumed sovereignty over Macao in December 1999 after it had been ruled since the mid-16th century by Portugal.<br><br>
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Antibiotics were added to each media (chloramphenicol 25mg/mL for Positive control、Negative control、Device 1：J23101+I13504、Device 2:J23106+I13504、Device 3:J23117+I13504). <br><br>
 
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Beijing is willing to carry out major projects with these countries by making use of the China-Portuguese-Speaking Countries Cooperation Development Fund and the Silk Road Fund, Li said.<br><br>
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We used100mL Luria Broth Liquid media(102μg/mL chloramphenicol) in 250 mL conical flask to grow our cells(37℃，110rmp). Every 1h, we took 100μL in 96-well plate (round wells, flat bottom) to test the OD600 and fluorescence intensity. <br><br>
 
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The announcement was made in Li's keynote speech on Tuesday at the opening ceremony of the two-day Fifth Ministerial Conference of the Forum for Economic and Trade Cooperation Between China and Portuguese-Speaking Countries.<br><br>
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Measurement of fluorescence and OD were performed using the Thermo Scientific Varioskan Flash with the 'Thermo Scientific SkanIt' software and with the 96-Well Multiwell Plates (Corning). <br><br>
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Delegates from seven Portuguese speaking countries — Portugal, Brazil, Cape Verde, Guinea-Bissau, Mozambique, Angola and East Timor — attended the forum.<br><br>
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Before the ceremony, Li launched a trade and economic cooperation program to promote further exchanges between China and Portuguese-speaking countries.
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China and the Portuguese-speaking countries contribute 17 percent of the world economy and 22 percent of global population, with varying advantages in technology, capital and market practices, Li said.<br><br>
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The participating countries should consolidate political trust, promote trade liberalization and investment and strengthen cooperation in production capacity and people-to-people exchanges, the premier told the forum.<br><br>
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China's economic restructuring and upgrading will bring great investment opportunities and market potential, creating bright prospects for businesses from Portuguese-speaking countries, Li said.<br><br>
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He said the trade volume between China and Portuguese-speaking countries reached $100 billion last year — about 10 times of that of 2003.<br><br>
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Cape Verde's Prime Minister Ulisses Correia e Silva said the Macao forum injected new momentum into strategic cooperation between all the participating countries. He added that his country will align with China's Belt and Road Initiative and make use of the Macao forum to bring more commodities to Africa.<br><br>
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The prime ministers of Guinea-Bissau and Mozambique vowed to adopt favorable policies and improve the rule of law to attract investors.<br><br>
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Deng Yanzi contributed to this story<br><br><br><br><br><br>
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<h2><br><br><br><br>Photometry</h2>
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Latest revision as of 03:33, 20 October 2016

<!DOCTYPE html> Biology

JUDGING

INTERLAB STUDY





Summary

The objective of this year’s iGEM Interlab study is to quantify expression of five different reporter constructs. These constructs have GFP under the control of different promoter and ribosome binding sequences (RBS). Our Interlab work was carried out with the DH5-alpha as an expression host, and the Thermo Scientific Varioskan Flash spectral scanning as the equipment. We transfer the five reporter constructs into E.coli cells separately. And we measure the fluorescence data and optical density data with spectral scanning machine every 1 hour during a 6 hours cultivation.





Methods

We use endonuclease (EcoRI and PstI, XbaI and PstI) and T4 DNA ligase (NEB) to construct them. Finally, the ligation product was introduced into the pSB1C3. All constructs used were transformed into DH5α cells.

The cultivation of our bacteria was performed in tubes filled with 7 mL LB liquid medium. The cultures were kept at 37 °C and 110 rpm in shaker.

Antibiotics were added to each media (chloramphenicol 25mg/mL for Positive control、Negative control、Device 1：J23101+I13504、Device 2:J23106+I13504、Device 3:J23117+I13504).

We used100mL Luria Broth Liquid media(102μg/mL chloramphenicol) in 250 mL conical flask to grow our cells(37℃，110rmp). Every 1h, we took 100μL in 96-well plate (round wells, flat bottom) to test the OD600 and fluorescence intensity.

Measurement of fluorescence and OD were performed using the Thermo Scientific Varioskan Flash with the 'Thermo Scientific SkanIt' software and with the 96-Well Multiwell Plates (Corning).





Photometry

...







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