Difference between revisions of "Team:IIT Kharagpur/Results"

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{{IIT_Kharagpur}}
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<html>
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<head>
  
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<title>IGEM-IIT Kharagpur- Part</title>
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<p>Here you can describe the results of your project and your future plans. </p>
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<h5>What should this page contain?</h5>
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<li> Clearly and objectively describe the results of your work.</li>
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    100%{ transform:rotate(360deg); }
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<li> Future plans for the project </li>
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}
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<li> Considerations for replicating the experiments </li>
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.icon{
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<div class="column half_size" >
 
  
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}
  
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<h5> Project Achievements </h5>
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  #logokgp{
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<p>You can also include a list of bullet points (and links) of the successes and failures you have had over your summer. It is a quick reference page for the judges to see what you achieved during your summer.</p>
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}
 +
}
  
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<ul>
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#dep{
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<li>A list of linked bullet points of the successful results during your project</li>
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<li>A list of linked bullet points of the unsuccessful results during your project. This is about being scientifically honest. If you worked on an area for a long time with no success, tell us so we know where you put your effort.</li>
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font-size: 1.4em;
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<h5>Inspiration</h5>
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h2 {
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<p>See how other teams presented their results.</p>
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font-size: 2em;
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<ul>
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line-height: 1.25em;
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<li><a href="https://2014.igem.org/Team:TU_Darmstadt/Results/Pathway">2014 TU Darmstadt </a></li>
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margin: .25em 0;
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<li><a href="https://2014.igem.org/Team:Imperial/Results">2014 Imperial </a></li>
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}
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<li><a href="https://2014.igem.org/Team:Paris_Bettencourt/Results">2014 Paris Bettencourt </a></li>
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</ul>
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<body >
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<!--=== Preloader section starts ===-->
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<span class="icon-bar"></span>
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<span class="icon-bar"></span>
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<!--=== Site Name ===-->
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<a class="site-name navbar-brand" href="https://2016.igem.org/Team:IIT_Kharagpur"><span>IIT</span>Kharagpur</a>
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<li class="current dropdown"><a href="https://2016.igem.org/Team:IIT_Kharagpur"><img src="https://static.igem.org/mediawiki/2016/1/13/T--IIT_Kharagpur--home_icon.png" class="icon"> Home</a>
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    <a href="https://2016.igem.org/Team:IIT_Kharagpur/Notebook" class="submenuhover">Notebook</a>
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<a href="https://2016.igem.org/Team:IIT_Kharagpur/Protocol" class="submenuhover">Protocol</a>
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    <a href="https://2016.igem.org/Team:IIT_Kharagpur/Data_page" class="submenuhover">Data Page</a>
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<a href="https://2016.igem.org/Team:IIT_Kharagpur/Safety" class="submenuhover">Safety</a>
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  </div>
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<a href="https://2016.igem.org/Team:IIT_Kharagpur/Collaborations" class="submenuhover">Collaboration</a>
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    <a href="https://2016.igem.org/Team:IIT_Kharagpur/Attributions" class="submenuhover">Attribution</a>
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<div class="dropdown-content">
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    <a href="#" class="submenuhover">Survey</a>
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<a href="#" class="submenuhover">Workshop</a>
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    <a href="#" class="submenuhover">Interview</a>
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<a href="#" class="submenuhover">Meet ups</a>
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  </div>
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</li>
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<li class="dropdown"><a href="https://2016.igem.org/Team:IIT_Kharagpur/Parts"><img src="https://static.igem.org/mediawiki/2016/7/76/T--IIT_Kharagpur--part_icon.png" class="icon">Part</a>
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    <a href="https://2016.igem.org/Team:IIT_Kharagpur/Parts" class="submenuhover">Composite parts</a>
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  </div>
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 +
<li><a href="blog.html"><img src="https://static.igem.org/mediawiki/2016/5/5e/T--IIT_Kharagpur--blog_icon.png" class="icon">Blog</a></li>
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<!-- sticky-bar Ends-->
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<!--=== Header section Ends ===-->
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<!--=== Home Section Starts ===-->
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<div class="topcontainer home">
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<div class="row">
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<div class="home_content">
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<h1 class="well-come" style="font-family:Dosis;">RESULTS</h1>
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<img src="https://static.igem.org/mediawiki/2016/7/76/T--IIT_Kharagpur--part_icon.png" class="center_icon">
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<!-- <div class="col-md-8 col-md-offset-2 tabledown">
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<p class="intro-message" style="font-family:Dosis; font-size:20px; text-align:center;">HARD WORK JUST LIKE JUST MA AND PA TAUGHT US</p>
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</div>
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<div class="col-md-12 bodysection">
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<div class="col-md-10 col-md-offset-1 center section-titler section-title">
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<h3>Overview</h3>
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</div>
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<div class="col-lg-12 col-md-12 col-sm-12 justify safety-content">
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<p style="font-family:Dosis; font-size:20px;">
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In an attempt to synthesize two constructs, one for the Silk and the other for the FRET, we managed to move only in the direction of the FRET construct. Owing to time restrictions along with unsatisfactory results, the silk construct could not be worked upon.
 +
For the FRET construct, we managed to construct the part in two halves. One of the parts, BBa_K2163000, comprises of Cyan Fluorescent Protein, HIV Protease Cleavage Site, and Yellow Fluorescent Protein. This composite part was prepared by standard assembly.
 +
The other half of the FRET construct consists of pBAD/AraC promoter, RBS, and the OmpA protein. We isolated this part from an existing construct using PCR amplification. However, we failed to insert the amplified product into a backbone and thus could not complete the entire part.
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</p>
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</div>
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</div>
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<div class="col-md-12 bodysection">
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<div class="col-md-10 col-md-offset-1 center section-titler section-title">
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<h3>RESULTS</h3>
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</div>
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<div class="col-lg-7 col-md-7 col-sm-12 justify safety-content">
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<h4><b>CFP+HIV Cleavage Site:
 +
 +
</b></h4>
 +
<p style="font-family:Dosis; font-size:20px;">
 +
The part was synthesized and checked using 1% agarose gel. Ligations were done in 2 sets, with the ratios 1:3 and 1:5, and 14 colonies from each were screened. A significant number showed the necessary shift.
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 +
</p><br><br><br>
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</div><br>
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<div class="col-lg-5 col-md-5 col-sm-12  justify safety-content" >
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<img src="https://static.igem.org/mediawiki/2016/7/7a/T--IIT_Kharagpur--27_Sep_C_H_Screen.jpg" height="400px;" width="400px;"><br><br><br>
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</div>
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<div class="col-lg-7 col-md-7 col-sm-12 justify safety-content">
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<p style="font-family:Dosis; font-size:20px;">
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Further, we digested the product with XbaI and PstI. We got the expected results after an agarose gel run.
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 +
</p>
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<br><br><br>
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</div><br>
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<div class="col-lg-5 col-md-5 col-sm-12  justify safety-content" >
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<img src="https://static.igem.org/mediawiki/2016/a/a7/T--IIT_Kharagpur--1_Oct_C_H_Digestion.jpg" height="400px;" width="400px;"><br><br><br>
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</div>
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<div class="col-lg-7 col-md-7 col-sm-12 justify safety-content">
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<h4><b>CFP+HIV Cleavage Site+YFP:
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 +
</b></h4>
 +
<p style="font-family:Dosis; font-size:20px;">
 +
The part was synthesized and checked using 1% agarose gel. Ligations were done in 2 sets, with the ratios 1:3 and 1:5, and 14 colonies from each were screened.
 +
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</p><br><br><br>
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</div><br>
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<div class="col-lg-5 col-md-5 col-sm-12  justify safety-content" >
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<img src="https://static.igem.org/mediawiki/2016/1/18/T--IIT_Kharagpur--6_Oct_C_H_Y_Screen.jpg" height="400px;" width="400px;"><br><br><br>
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</div>
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<div class="col-lg-7 col-md-7 col-sm-12 justify safety-content">
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<p style="font-family:Dosis; font-size:20px;">
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Further, we digested the product with XbaI and PstI. We got the expected results after an agarose gel run.
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<br><br><br>
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</div><br>
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<div class="col-lg-5 col-md-5 col-sm-12  justify safety-content" >
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<img src="https://static.igem.org/mediawiki/2016/5/52/T--IIT_Kharagpur--7_Oct_C_H_Y_Digestion.jpg" height="400px;" width="400px;"><br><br><br>
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</div>
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<div class="col-lg-7 col-md-7 col-sm-12 justify safety-content">
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<h4><b>Promoter + RBS + OmpA:
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 +
</b></h4>
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<p style="font-family:Dosis; font-size:20px;">
 +
We amplified the part by design suitable primers for BBa_K43013, the protease construct designed by Team Chile. We were able to standardize the PCR conditions, but failed at insertion into a backbone.
 +
 +
</p>
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<br><br><br>
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</div><br>
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<div class="col-lg-5 col-md-5 col-sm-12  justify safety-content" >
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<img src="https://static.igem.org/mediawiki/2016/0/08/T--IIT_Kharagpur--8_Sep_fret_pcr.jpg" height="400px;" width="400px;"><br><br><br>
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</div>
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<div class="col-lg-12 col-md-12 col-sm-12 justify safety-content">
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<h4><b>Further  Possibilities:
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 +
 +
</b></h4>
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<p style="font-family:Dosis; font-size:20px;" style="margin-bottom:30px;">Through this year’s project, we realised the potential of the FRET system. And accordingly, we will continue to work on the same, and try to find application in various areas.
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We were disheartened that we could not crack the Silk construct. Although, we will continue work on this construct too, and hope to complete the same. The silk construct require more modification to enhance efficiency. We plan to tackle the problem as we go along.
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Latest revision as of 18:44, 19 October 2016

IGEM-IIT Kharagpur- Part

Overview

In an attempt to synthesize two constructs, one for the Silk and the other for the FRET, we managed to move only in the direction of the FRET construct. Owing to time restrictions along with unsatisfactory results, the silk construct could not be worked upon. For the FRET construct, we managed to construct the part in two halves. One of the parts, BBa_K2163000, comprises of Cyan Fluorescent Protein, HIV Protease Cleavage Site, and Yellow Fluorescent Protein. This composite part was prepared by standard assembly. The other half of the FRET construct consists of pBAD/AraC promoter, RBS, and the OmpA protein. We isolated this part from an existing construct using PCR amplification. However, we failed to insert the amplified product into a backbone and thus could not complete the entire part.

RESULTS

CFP+HIV Cleavage Site:

The part was synthesized and checked using 1% agarose gel. Ligations were done in 2 sets, with the ratios 1:3 and 1:5, and 14 colonies from each were screened. A significant number showed the necessary shift.








Further, we digested the product with XbaI and PstI. We got the expected results after an agarose gel run.








CFP+HIV Cleavage Site+YFP:

The part was synthesized and checked using 1% agarose gel. Ligations were done in 2 sets, with the ratios 1:3 and 1:5, and 14 colonies from each were screened.








Further, we digested the product with XbaI and PstI. We got the expected results after an agarose gel run.






Promoter + RBS + OmpA:

We amplified the part by design suitable primers for BBa_K43013, the protease construct designed by Team Chile. We were able to standardize the PCR conditions, but failed at insertion into a backbone.








Further Possibilities:

Through this year’s project, we realised the potential of the FRET system. And accordingly, we will continue to work on the same, and try to find application in various areas. We were disheartened that we could not crack the Silk construct. Although, we will continue work on this construct too, and hope to complete the same. The silk construct require more modification to enhance efficiency. We plan to tackle the problem as we go along.